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【Objective】The objective of this study is to identify and reveal the transition of the male sterile gene(s) in photoperiod- and temperature-sensitive genic male sterile (P/TGMS) lines utilized in the two-line hybrid rice system in China.【Method】A total of 90 environment-conditioned genic male sterile (EGMS) lines including descendents of Nonken 58S, Annong S-1 and Zhu 1S, were used in the present study. Genomic DNAs were extracted from rice leaves by modified CTAB. One functional CAPS marker based on the C to G mutation in the long non-coding RNA (lncR) gene was designed for PGMS genotyping; Namely, a pair of primers NK-F (5′-ATCCCACAAATCCTTTAGCA-3′) and NK-R (5′-CCGTTATAGATAGACCCGAGA-3′) were used to amplify segments harboring the mutation site, followed by digestion overnight at 37℃ with restriction endonuclease RsaⅠ and separation on 1% agarose gel electrophoresis. Homozygous PGMS allele (lncR^m) (329 bp) can be readily distinguished from homozygous wild type (lncR^wt) (414 bp) and heterozygous type (lncR^m/lncR^wt) (414 and 329 bp) based on the sizes of digestion products. For TGMS genotyping, functional dCAPS markers were deployed with the following steps: Two pairs of primers RNZ1F (5′-ACCGCGCCGCCACCGGGTCGGCCGGAG-3′)/RNZR (5′-TGAAGAGGAACTCCTGCGAGACGG-3′), RNZ2F (5′-ACCGC GCCGCCACCGGGTCGGCCCAAG-3′)/RNZR were used to amplify segments harboring the mutation site (SNP-^+70TA/TC/GC); Amplified products were digested overnight at 37℃ with restriction endonucleases HinfⅠand StyⅠ, respectively, and separated on 8% polyacrylamide gels. Homozygous lines with the TGMS allele (RNZ^m) cannot be digested by these two restriction enzymes; on the other hand, homozygous wild type lines with the alleles of RNZ^tc or RNZ^gc, can be digested completely by HinfⅠ and StyⅠ, respectively; heterozygous genotypes, RNZ^m/RNZ^tc and RNZ^m/RNZ^gc, can be digested incompletely by HinfⅠand StyⅠ, respectively. By using these functional molecular markers, the PGMS (lncR^m) and TGMS (RNZ^m) genes were identified in commercial EGMS lines that had been utilized in the two-line hybrid rice system. Meanwhile, the transition of P/TGMS genes utilized in two-line hybrid rice production in China (1993-2012) was analyzed according to the information of pedigree and growing area. 【Result】Out of the 47 EGMS lines derived from the PGMS line Nongken 58S, 12 lines carry the PGMS gene lncR^m, 29 lines have the TGMS gene RNZ^m, two lines carry both genes, while the remaining four lines contain none of them. All 18 lines derived from Annong S-1 and Zhu 1S carry RNZ^m gene. All P/TGMS descendants from crosses between a Nongken 58S derivative (Pei’ai 64S) and Annong S-1 carry RNZ^m gene. In the two lines derived from Pei’ai 64S and Zhu 1S, one carries both genes, the other has RNZ^m gene. In addition, in 16 EGMS lines with EGMS progenitors independent from Nongken 58S, Annong S-1 and Zhu 1S, six of them have lncR^m, nine of them have RNZ^m gene, while one of them contains neither lncR^m nor RNZ^m gene. A pedigree map with P/TGMS genes was drawn for 92 EGMS lines, including derivatives from Nongken 58S, Annong S-1, Zhu 1S and others. Furthermore, the transition from lncR^m-based to RNZ^m-based two-line hybrid rice production was shown after examination of statistics data of the two-line hybrid rice planting area during 1993-2012, with the RNZ^m-based hybrids occupying ＞95% planting area in the two-line hybrid rice production in 2012. 【Conclusion】The study systematically revealed the P/TGMS genes in commercial EGMS rice and the presence of discrepancy between pedigree of EGMS lines and their P/TGMS gene. The spontaneous emergence of RNZ^m can be the reason for the transition from PGMS to TGMS in some EGMS lines derived from Nongken 58S. EGMS lines with RNZ^m currently dominate the two-line hybrid rice production in China.

【Objective】 The present study was aimed to take a first step of the improvement of the previously reported chromosome segment substitution line (CSSL) population SojaCSSLP1, and to explore superior QTL/gene-alleles related to some agronomic traits from the wild parent (Glycine soja Sieb. et Zucc.) for broadening the genetic basis of cultivated soybean (Glycine max (L.) Merr.). 【Method】 The SojaCSSLP1, with the wild soybean N24852 as donor parent and the cultivated soybean NN1138-2 as recurrent parent, was treated with adding markers and removing a number of lines with segment of single marker, the new population was designated as SojaCSSLP2. By using the new population, the QTL/segments for flowering time (FT), plant height (PH), node number (NN), pod number per plant (PN), 100-seed weight (100SW) and seed weight per plant (PSW) were detected through joint comparisons among CSSLs significantly different from the recurrent parent based on QTL mapping with the methods of single marker analysis (SMA), interval mapping (IM), inclusive composite interval mapping (ICIM) and mixed linear composite interval mapping (MCIM), for experiments in three years each with two locations. 【Result】SojaCSSLP2 was composed of 150 CSSLs, of which 130 ones were the same as SojaCSSLP1, added 40 new SSR markers into the previous molecular map, resulted in the average genetic distance and the number of lines with genetic distance more than 30 cM between adjacent markers reduced from 16.15 cM and 32 to 12.91 cM and 17, respectively. The total length of the genetic map increased by 103.52 cM in comparison to the original map (2 063.04 cM). The genome component of NN1138-2 in CSSLs ranged from 79.45% to 99.70% with an average of 94.62% in SojaCSSLP2. Tested in three years and two locations, 4, 5, 5, 7, 14 and 3 working QTL/segments were identified for FT, PH, NN, PN, 100SW and PSW, respectively, with the improved population. Of those, 15 QTL/segments were joint working QTL which could be detected in more than one environment. The directions of additive effects for wild alleles were consistent with that expected from the parental phenotypes except segment of Sct_190-Sat_293 for NN. Among the QTL, the individual QTL could explain 5% to 64% of the phenotypic variation. There were 3, 2 and 2 QTL/fragments that interacted with locations for PH, NN and NP, respectively, and the interactions associated with Fengyang could increase the value of phenotype, which may be due to its higher latitude than that of Nanjing. These detected loci distributed on 26 substituted segments, among them seven related to more than one trait, which might be the genetic basis of correlation among the traits. Compared with the results in the literature, 3, 3, 2, 2, 8 and 2 QTL could also be detected in other cultivated soybeans for FT, PH, NN, 100SW and PSW, respectively, indicating allele differentiation happened not only between wild and cultivated but also among cultivated soybeans. The other 18 loci/segments were newly discovered in the wild soybean.【Conclusion】 The genetic base of FT, PH and NN are much simpler than that of 100SW. There was a large effect QTL (PV＜10%) in the former, while the later is controlled by many small effect loci (PV＜10%) with complicated genetic bases. Using the wild soybean, the novel alleles with the capability of broadening the genetic base of cultivated soybean can be explored.

【Objective】 Soybean seed is susceptible to high temperature and humidity (HTH) stress resulting in pre-harvest deterioration during its development and maturation (R6 or R7), which limits the production and utilization of soybean in south China. The aim of the present study is to reveal the pre-harvest seed deterioration and deterioration resistance mechanism of spring soybean under HTH stress at proteomic level, thus laying a foundation for breeding new cultivars.【Method】When the deterioration-sensitive cultivar Ningzhen No.1 and deterioration-tolerant cultivar Xiangdou No.3 developed to physiological period (R7), their plants were treated under HTH stress and control for 1, 5, 10, 16 and 24 h, respectively. The extracted proteins from the developing seed of treatment and control were analyzed by two-dimensional electrophoresis (2-DE), and the differentially expressed proteins were identified by MALDI-TOF/TOF. 【Result】Approximately 700 protein spots were detected on each 2-DE gel. Of them, 50 were found to be significantly changed in abundance, with 33 being successfully identified by MALDI-TOF/TOF. The identified differentially expressed proteins were involved in cell rescue and defense (9%), redox homeostasis (12%), protein synthesis (3%), energy metabolism (15%), transport pathway (15%), protein destination and storage (31%). Moreover, the function of 5 identified proteins is unknown. 【Conclusion】Pre-harvest deterioration-resistant cultivar Xiangdou No.3 possessed the greater ability of ROS scavenging and cell rescue and defense than deterioration-sensitive cultivar Ningzhen No.1 under HTH tress, which may be the major reasons why it is more deterioration-resistant than the latter.

【Objective】This experiment was carried out to understand the viability and physiological variation rule of different genotypes maize seeds under critical stress storage conditions, evaluate its of storability objectivity, provide a theoretical basis for clearing the physiological and biochemical mechanisms of maize seed storability. 【Method】 Major maize hybrids(ZD958, ND108, XY335, ZD619, JK968) were used as experimental materials. Seed initial moisture content, vigor and physiological indicators was measured. The moisture content of experimental materials wet back to 14% and stored at temperature of 35℃, for one year, then the germination energy, germination rate, germination index, vigor index, electrical conductivity, MDA concentration, soluble protein and soluble sugar content and dehydrogenase activity of materials were measured every month. The germination experiment was conducted by using sand culture method, DDS-ⅡA conductivity meter was used to measure the seed electrical conductivity, the TBA was used to measure the MDA content, coomassie brilliant blue G-250 method was used to analyze the soluble sugar content, and dehydrogenase activity was determined by TTC method. The vigor and physiological indicators change of different genotypes maize seeds were compared under the critical stress storage conditions.【Result】Seed genotype was the determinative factor under the same storage condition. The initial moisture content of experimental materials was at a low level, between 7.39% and 8.71%. The germination potential and germination rate of different genotypes maize seed were over 90%, showing a stronger germination ability. After storage for one year, the germination potential and germination rate of JK968 was 50%-60%, germination index was 25%, and vigor index was 0.3. The germination potential and germination rate of ND108, XY335, ZD619 was 15%-25%, germination index was 8%-25%, and vigor index was 0-0.08. The germination potential, germination rate, germination index and vigor index of ZD958 decreased to 0 after storage for one year. Different degrees in decline of germination potential, seed soluble protein content and soluble sugar content changes showed a close relationship with seed vigor. Germination rate, germination index, vigor index were almost the same, however, there was a number of differences in amplitude of variation. Different germination speeds lead to different germination indexes of each genotype of maize seed, seed vigor index declined was always ahead of seed germination ability, which reflected the degree of real seed aging and deterioration. With the storage time extension, the membrane permeability variation of different genotypes of maize seeds, electrical conductivity and MDA content were increased and the change trend was negatively related with seed vigor(R²=0.752). A membrane permeability difference existed among different genotypes of maize seeds. The vigor of different genotypes of maize seed had no significant correlation with the MDA(R²=-0.171-0.094), it significantly positively correlated with soluble protein, soluble sugar and dehydrogenase (R²=0.284- 0.517), but the mechanism of physiological changes of different genotypes of maize seed was complex, the gap was bigger.【Conclusion】 Different genotypes of maize seed were more sensitive to critical stress storage condition, ZD958 was sensitive to critical stress, the vigor and physiological indicator changes of JK968 were stable, showed a higher storability.

【Objective】 Intercropping, as one of the most universal multiple cropping systems based on biodiversity, plays an important role in enhancing crop productivity for more efficient use of resources, such as land, light, temperature and water as well as reducing incidence of weeds, insects, pests and diseases to meet the future human demand. Cereal-legume relay intercropping system is commonly used in China, because of the different distances between maize and soybean which influence the PAR distribution above the soybean canopy within relay strip intercropping system, and it is not only bad to the development of canopy structure but also decreases the light interception of intercropped soybean, thus resulting in limited photosynthetic active radiation use efficiency of intercropped soybean which restricts the productivity of maize-soybean relay strip intercropping systems. Therefore, there is a need for exploring an optimum spatial-temporal configuration management of intercropping system based on competition so that light resources requirement of soybean can be realized in this system. 【Method】 An experiment with maize (Zea mays L.) cultivar Chuandan 418 and soybean (Glycine max L. Merr.) cultivar Gongxuan1 were used in this study. The relay strip intercropping systems were designed as the distances of maize intercropped with soybean were 40 cm (A)，50 cm (B) and 60 cm (C), and both of maize monoculture (SM) and soybean monoculture (SSB) were used as control. The soybean population canopy structure, light interception, dry matter weight in the relay strip intercropping combinations were measured and analyzed under maize/soybean relay strip intercropping systems in order to optimize the reasonable group configuration.【Result】 The PAR density was different above the soybean canopy under different maize-soybean relay strip intercropping systems, and lower than SSB significantly (P＜0.05). During the co-growth stage of maize and soybean, the PAR density occurred in treatment A was lower than that in treatments B and C by 44.1% and 60.4%, respectively. This means that the PAR density decreased as the degree of low light stress increasing due to the distance between maize and soybean decreased. The LAI, LA and plant height of intercropped soybean varied considerably in different maize-soybean relay strip intercropping systems. During the stages of V5, V7 and R1, the LAI of treatment B was higher than treatment A and treatment C significantly. Treatment B was higher than that in treatments C and A by 16.4%, 13.1%, 12% and 30.3%, 32.2%, 29.3%. LA of treatment B was higher than that in treatments C and A by 15%, 16%, 14% and 34%，31%，26%. Plant height of treatment B was lower than that in treatments C and A by 7%, 8.8%, 7.9% and 13.5%, 16.7%, 14.8%, which means that suitable soybean canopy structure can improve RUE. The LI of intercropped soybean was decreased significantly under different maize-soybean relay strip intercropping. During the stages of V5, V7 and R1, the LI of treatments A and C was lower than that in treatment B by 43%, 22%, 33% and 21%, 10%, 17%. A positive correlation of LAI and LI was significant (0.977**), which means that LAI might be the main factor due to the LI increasing. The photosynthetic active radiation use efficiency of intercropped soybean varied considerably in different maize-soybean relay strip intercropping systems. During the stages of V5, V7 and R1, the RUE of treatment B was higher than that in treatments C and A by 8.6%, 7%, 5.8% and 40%, 23%, 13%. The dry matter weight of soybean in all intercropping systems was lower than monoculture significantly (P＜0.05), and varied considerably in different maize and soybean relay strip intercropping systems. During the stages of V5, V7 and R1, the dry matter weight of treatments A and C was lower than that in treatment B by 59%, 36%, 41% and 27%, 16% and 22%. A positive correlation of DMW and LI was significant (0.989**), which means that the dry matter increased with the light interception increasing. The yield of intercropped soybean and total yield varied considerably in different maize and soybean relay strip intercropping systems(P＜0.05). The yield of treatment B was higher than that in treatments C and A by 10% and 27%, the total yield of treatment B was higher than that in treatments C and A by 1% and 3%. This means that the yield of intercropped soybean increased/decreased as influenced by the intensified low light stress due to the difference of distance between maize and soybean. 【Conclusion】In this case, suitable distance of maize and soybean configuration could optimize intercropped soybean canopy structure, improve the light use efficiency and enhance yield.

【Objective】Wheat sharp eyespot (WSE) caused by Rhizoctonia cerealis is one of the most important soil-born diseases on wheat in China. Early accurate quantitative detection is a foundation of forecast and control. Traditional method of organization isolation and identification of pathogen is time consuming, complicated and can’t be accurately quantified. In order to implement the early and quick quantitative determination of wheat sharp eyespot, a SYBR Green I real-time PCR method of R. cerealis was established based on the pathogen sequence information.【Method】 Based on the β-tubilin of R. cerealis, a pair of specific primers was designed. The SYBR Green I real-time PCR reaction system was established and optimized. The sensitivity, specificity and repeatability of the system were also evaluated, and RBipolaris sorokiniana, Fusarium graminearum, F. pseudograminearum were used for control fungi. The indoor potted plants of wheat which infected by R. cerealis were detected with optimized reaction system after inoculated for 5, 10 and 60 days, respectively.【Result】The primers were of great specificity, the specific PCR fragment was amplified from the DNA of R. cerealis isolates, but not from the DNA of other fungal isolates by conventional PCR. The real-time PCR assays also did not amplify DNA from control fungi. The sensitivity of conventional PCR was 6.5×10³ copies/μL plasmid, while the sensitivity of real-time PCR was 6.5×10² copies/μL. The standard curve established by recombinant plasmid showed a fine linear relationship between threshold cycle and template concentration. The melt curve was specific with the correlation coefficient of 0.997 and with high amplification efficiency (0.91). For the indoor potted experiments, the detection results of real-time PCR of infected wheat samples, were showed a significant positive correlation with disease index and inoculum, respectively. 【Conclusion】 The developed real-time PCR assay for R. cerealis is fast, highly specific, sensitive, and reproducible. This method can be used to detect R. cerealis in wheat, and guidance prediction and control of wheat sharp eyespot.. cerealis, R. solani, AG-A, AG-F, Gaeumannomyces graminis var. tritici,

【Objective】The objective of this experiment is to study the effect of Huanglongbing (HLB) infection on carbohydrate metabolism in Citrus sinensis, elucidate the relationship between HLB infection and starch accumulation, and provide a theoretical basis for HLB pathogenesis. 【Method】C. sinensis were used as the experimental materials, plants were inoculated by grafting three bark pieces onto the rootstock portion of each plant. For infected plants, bark pieces were derived from trees infected with Candidatus Liberibacter asiaticus, which were confirmed positive for Ca. L. asiaticus by PCR. For non-infected plants, bark pieces were obtained from healthy trees. Three plants in good growth state were chosen for successive test, mature leaves were collected every month until October. DNA and RNA were extracted, the content of soluble sugar and starch were measured immediately, the other leaf tissue was snap-frozen with liquid nitrogen and stored at -80℃ for testing carbohydrate metabolism related key enzyme activities. At the same time, the expression levels were compared using real-time quantitative PCR analysis. 【Result】The content of soluble sugar and starch tended to increase with time extended, and their peak value were respectively reached 1.59 and 3.73 times of control in June, then slightly decreased later, the ratio of soluble sugar to starch dropped down. Various key enzymes imposed different hands on carbohydrate metabolism at different stages of infection. Sucrose phosphate synthase activity rapidly rose to the top 58.44 μg?g^-1?min^-1 and had become lower than control in late infection; acid invertase had been consistently high and the difference was significant (P＜0.05), the highest activity was 2.91 times of control at middle stage; the activity of neutral invertase was kept at a lower level and had the same change trend in experimental group and control group; soluble starch synthase activity was closely related to granule-bond starch synthase, coordinating the synthesis of acting on the starch, while amylase activity had dropped to some extend at each stage, even as low as 0.38 U?g^-1 at middle stage. Real-time quantitative PCR indicated that there was a significant relationship between the expression level of sucrose phosphate synthase and its enzyme activity (P＜0.05). Among the sucrose decomposition related genes, sucrose synthase gene was not changed obviously and had lower change range while cell wall-bound invertase gene CSCWI was up-regulated with the highest value of about 7.4 times and expressed abundantly during the whole process. Starch decomposition-related genes BAM3, MEX1 and DPE2 had a relatively reduced expression and the difference was significant (P＜0.05) in different periods.【Conclusion】HLB infection has influence on the formation and transportation of photosynthate in C. sinensis and results in the disruption of carbohydrate metabolism balance, it is in deep connection with starch accumulation and the late stage symptoms.

【Objective】Based on the statistic data of cultivated area, sown area, crop yield, production value and farmland investment of the crop production in the period of 1993 to 2012, the dynamics, distributions and compositions of carbon footprint (CF) of the crop production were estimated in North China Plain (NCP), which will provide a theoretical support and scientific basis for the development of low-carbon agriculture. 【Method】Based on the historical statistic data of cultivated area, sown area, crop yield, production value and farmland investment of the crop production of 47 prefecture-level cities in 5 provinces and cities (Hebei, Beijing, Tianjin, Shandong and Henan) in North China Plain, and used the theory of carbon footprint and life cycle assessment method in the agricultural sector, CF per unit sown area (CFs), CF per unit cultivated area (CFc), CF per unit yield (CFy) and CF per unit production value (CFv) of the crop production were estimated in North China Plain. The dynamic and distributions of carbon footprint were analyzed briefly. In addition, the proportions of different agricultural inputs in carbon footprint of crop production were explored.【Result】 CFs and CFc of crop production showed an increasing trend in the last twenty years. The CFs of crop production was 4.08 tCO₂eq·ha^-1 in period I (1993-2002) and 4.72 tCO₂eq·ha^-1in period II (2003-2012), respectively; The CFc of crop production was 6.81 tCO₂eq·ha^-1 in period I (1993-2002) and 8.12 tCO₂eq·ha^-1in period II (2003-2012), respectively. CFy and CFv of crop production showed a decreasing trend in the last twenty years. The CFy of crop production was 0.54 tCO₂eq·t^-1 in period I and 0.42 tCO₂eq·t^-1in period II, respectively. The CFv of crop production was 3.91 tCO₂eq·¥10^-4 in period I and 2.51 tCO₂eq·¥10^-4 in period II, respectively. The carbon footprints were different in different regions of the NCP. Most coastal regions (Tangshan, Tianjin and Yantai, etc.) have bigger CFs and CFc and smaller CFy and CFv. On the contrary, most cities of South Henan (Zhumadian, Xinyang, and Pingdingshan, etc.) have smaller CFs and CFc and bigger CFy and CFv. The agricultural inputs per unit sown area and the vegetable crops sown area percentage in high yield region were significantly higher than that in low yield region, respectively. The results presented here indicated that the main components of carbon footprint were electricity for irrigation (30.25%), N fertilizer (23.07%), and direct emissions of N₂O (19.83%).【Conclusion】In the last twenty years, the CFs and CFc of crop production increased, but the CFy and CFvdecreased in North China Plain. Most coastal regions have bigger CFs and CFc and smaller CFy and CFv, but the regions of southern Henan province were the opposite. In addition, the electricity for irrigation and N fertilizer were the major resources of emissions in North China Plain.

Livestock production is one of the important emission sources of greenhouse gases (GHGs), evaluation of the carbon footprint of livestock products is vital for selection of mitigation technology and promotion of low-carbon agriculture. Based on current evaluation methods of carbon footprint, this study summarized the domestic and overseas researches on assessment of the carbon footprint of animal products (eggs, pork, beef and milk), and made a comprehensive analysis based on the present research achievements. Carbon footprint of livestock products varies with unit of animal products. The carbon footprint in producing 1 kg of beef is the greatest and reaches (20.51±8.39) kg CO₂-eq, followed by 1 kg of pork and eggs production with (4.24±1.07) kg CO₂-eq and (2.24±0.83) kg CO₂-eq, respectively, while that in producing 1 kg milk is the minimum of (1.19±0.40) kg CO₂-eq. The carbon footprint in producing 1 kg protein from animal products is in a descending order as beef＞milk＞pork and egg, with values of (103.05±42.14), (39.72±13.20), (32.09±8.14) and (19.37±7.15) kg CO₂-eq, respectively. The carbon footprint in producing 1 kg fat from animal products is in a descending order as beef＞milk＞egg and pork, with values of (488.25±199.65), (37.23±12.37), (29.28±10.80) and (11.45±2.91) kg CO₂-eq, respectively. The carbon footprint of 1 000 kcal energy from animal products are beef, milk, egg and pork in descending order, with values of (16.41±6.71), (2.21±0.73), (1.56±0.57) and (1.07±0.27) kg CO₂-eq, respectively. Analyses on different links of animal products revealed that the share of greenhouse gas emissions reaches maximum in feed crop planting and producing & processing during egg and pork production, accounting for (74.0±16.5)% and (61.3±7.6)%, respectively. The methane emission from enteric fermentation delivers the greatest contribution to carbon footprint during beef and milk production, accounting for (53.7±8.2)% and (52.7±6.1)%, respectively. Analyses on GHGs emissions from animal products showed that CO₂ contributes the maximum to the carbon footprint during egg production, whose emission covers (55.42±2.7)% of the entire system. N₂O contributes the maximum during pork production, with (56.8±10.4)% of the entire system. CH₄ contributes the maximum during beef and milk production, with (50.2±8.3)% and (58.6±8.3)%, respectively. Although researches on carbon footprint of livestock products based on different methodologies are mostly in abroad, unified assessment guidance needs to be developed to evaluate the carbon footprint of livestock products. There is little research on carbon footprint in China, so it’s suggested that an assessment method suitable for actual production in China should be established. The result of this study could provide some preliminary data to the assessment of carbon footprint of livestock product and identification of mitigation options.

【Objective】Cucumber is one of the ten vegetables in the world and parhenocarpy is an important trait closely related to production and quality of cucumber. To explore the inheritance and QTL mapping for parthenocarpy in cucumber could provide a preliminary basis for further study on mechanism of parthenocarpy and molecular assistant selection breeding, and lay a theoretical foundation for breeding of parthenocarpy.【Method】In this study, The authers clipped eight female flowers on main stem and branches, respectively, for every individual plant and investigated parthenocarpic fruit once when all plants treament finished 8-10 days later to calculate the parthenocarpy percentage (numbers of parthenocarpis fruit/numbers of clipped female flower) in order to evaluate parthenocarpy ability. Two F₂ progenies derived from two crosses between EC1, a gynoecious parthenocarpic line, and two monoecious non-parthenocarpic lines 8419 and 14519 were constructed to determine the inheritance of parthenocarpy in cucumber. A linkage map from part of F₂ plants from the cross of EC1×8419 was constructed with JoinMap4.0 software by screening 1 335 SSR from 9930 and gy14 cucumber genome sequencing and 143 Indel primers from two parents resequencing and QTL detection for parthenocarpy was conducted with WinQTLcart2.5 software using F_2:3 families from the same cross. The candidate genes in major QTL region were predicted using bioinformatic analysis method.【Result】Parthenocarpy in EC1 was inherited quantitatively but segregated towards different parents in two F₂ progenies. A linkage map containing 7 chromosomes, 116 SSR and 9 Indel markers was constructed, which total length was 802.9 cM and average distance between two markers was 6.3 cM. QTL analysis identified 7 QTLs, Parth1, Parth2-1, Parth2-2, Parth3-1, Parth3-2, Parth5, and Parth7，distributing on chromosomes 1, 2, 3, 5, and 7. The major QTL Parth2-1 locating between SSR00684-SSR22083 was the only locus detected in two seasons, having two LOD scores of 9.0, 6.2 and R² of 17.4%, 10.2%, respectively, and its genetic and physical distance was 17.1 cM and 2.9 Mb. There were 307 genes in this region and two gene among them, Csa2M035330.1 and Csa2M070880.1, involved in plant hormone signal transduction would be candidate genes closely related to parthenocarpy, the rest were minor QTLs.【Conclusion】The inheritance of pathenocarpy was quantitative. The QTL Parth2-1 locating on chromosome 2 was the major QTL controlling parthenocarpy in cucumber and two genes in plant hormone pathway would be candidate genes. The results in this study would lay a foundation for fine mapping and gene cloning of major QTL of parthenocarpy in cucumber and for use in MAS breeding.

【Objective】Ginger is a thermophilic vegetable crop, but not tolerate high temperature. It is easily damaged under high temperature. To study the effect of exogenous spermidine on endogenous hormones and chloroplasts, the relationship among exogenous spermidine, endogenous hormones, and the chloroplast ultrastructure of ginger under heat stress was investigated.【Method】Laiwu Big Ginger was sandy cultured in a climate chamber with 12 h/12 h photoperiod under 28℃/18℃ (normal) and 38℃/28℃ (heat stress) conditions. Ginger root was treated with 0.5 mmol·L^-1spermidine. Relative water content, chlorophyll concentration, malondialdehyde content, electrolyte leakage, chlorophyll fluorescence parameters, reactive oxygen content and endogenous hormones metabolism of ginger leaves were investigated on 0, 5, 10, 15 and 20 d after treatment, moreover, the ultrastructures of chloroplasts and thylakoid were observed on 20 d after treatment. 【Result】Under heat stress, chloroplasts and thylakoid of ginger leaves were seriously damaged, and chlorophyll concentration significantly decreased with continuing stress. Chlorophyll fluorescence parameters including F_v/F_m, Ф_PSII, F_v′/F_m′, q_P and P decreased, and NPQ, β/α-1 and D increased, which mainly showed that its photochemical activity of PSII was decreased, so the  and H₂O₂ accumulated extensively resulted by excess energy, and then the MDA content and electrolyte leakage increased. Under heat stress, superoxide dismutase activity, abscisic acid and proline contents were significantly accumulated and then decreased in different treatment times. Ascorbate peroxidase activity, cytokinin and kinetin content kept a decreasing trend with continuing stress. However, relative water content, chlorophyll concentration, malondialdehyde content and electrolyte leakage of ginger leaves were recovered by exogenous application of spermidine. Besides, it also maintained the integrity of chloroplasts and thylakoid, regulated chlorophyll fluorescence parameters to normale, increased antioxidant enzymes activity and endogenous hormones concentration, and reduced the reactive oxygen content. 【Conclusion】Heat stress can damage ginger leaves, leading to function of chloroplasts and PSII disorder, and reactive oxygen and endogenous hormones abnormity. Exogenous application of spermidine can effectively reduce the damages of ginger caused by heat stress and improve its tolerance to heat stress, involving maintain endogenous hormones regular metabolism and chloroplasts normal physiological function.

【Objective】 The influencing mechanisms of orchard mulching grass on the photosynthetic apparatus as well as the physiological function of apple leaves in the northern of the Bohai Gulf were studied in order to provide a theoretical basis for organic cover measures in orchard. 【Method】 ‘Hanfu’ apple (Malus domestica Borkh./GM256/M.baccata Borkh.) and Digitaria sanguinalis (L.) Scop. were used as materials, a potted experiment was conducted to determine the effects of orchard mulching grass together with using N on microstructure, photosynthetic pigment contents, gas exchange parameters and overall function of photosynthetic apparatus of apple leaves. The experiment were divided into four parts which were mulching grass treatment (C₁N₀, 1 kg grass per pot), nitrogen fertilizer (C₀N₁, 3.4 g N per pot) and mulching grass + nitrogen fertilizer treatment (C₁N₁, 1 kg grass and 3.4 g N per pot) and control (CK). 【Result】 The results showed that C₁N₀ and C₀N₁ treatments increased the palisade tissue thickness as well as the ratio of PT/ST, however, compared with the control, there was no obvious difference in spongy tissue and the leaf thickness. Leaf microstructure showed that C₁N₁ treatment increased the thickness of the palisade tissue, spongy tissue and leaf by 8.45%, 12.91%, and 19.34%, respectively. It also significantly improved the ratio of PT/ST. Straw mulching treatment could change photosynthetic pigment contents, the ratio of chlorophyll and photosynthetic gas exchange parameters, among them, chlorophyll a content, total chlorophyll content, the ratio of chlorophyll a/b and net photosynthetic rate (P_n) increased by 22.7%, 12.71%, 23.42% and 22.83%, respectively. The contents of chlorophyll a, chlorophyll b and total chlorophyll in C₁N₁ treatmentwere 1.42, 1.04 and 1.37 times than that of control, it also increased the net photosynthetic rate (P_n) and water use efficiency (WUE) by 41.71% and 21.99%, respectively. Photosynthetic pigment content and photosynthetic physiological parameters were higher in C₀N₁ and C₁N₁ treatments than that of C₁N₀ treatment, and control was the lowest. The chlorophyll a fluorescence transient and 820 nm reflection kinetics of apple leaves changed obviously. The JIP-test showed that different treatments increased the chlorophyll fluorescence parameter including maximal photochemistry efficiency of PS Ⅱ (F_v/F_m), efficiency/probability that an electron moves further than Q_A^- (ψ_o), quantum yield for electron transport (φ_Eo), which showed C₁N₁＞C₀N₁＞C₁N₀＞CK. Photosynthetic performance index (PI_ABS), biggest redox ability (ΔI/I_o) of PSⅠ were significantly higher than that of control. Different kinds of treatments promoted the development of apple plant. There was no significant difference between C₁N₀ treatment and control in leaf length. However, C₀N₁ treatment increased leaf length by 10.63% compared with the control. C₁N₀ and C₀N₁ treatments not only increased leaf width and leaf area, but also promoted the development of the trunk circumference by 8.82% and 12.35%. In addition, apple leaf length, leaf width, leaf area and trunk circumference of C₁N₁ were 1.14, 1.19, 1.44 and 1.21 times higher than that of control, respectively. 【Conclusion】 Under potted experiment, orchard mulching grass and nitrogen fertilizer treatment could improve the overall function of photosynthetic apparatus, and the development level of apple plant significantly.

【Objective】 The aim of this study was to explore the relationships between GHSR and GHRL gene and body weight & size traits, and search the molecular genetic markers related to goat growth traits.【Method】Guizhou white goat and Guizhou black goat were chosen as subjects, the DNA pooling was constructed and the technology of direct sequencing of PCR products and PCR-SSCP were used to detect the single nucleotide polymorphism of GHSR and GHRL genes, and its association of polymorphisms and different genotypes and combined genotypes with growth traits was analyzed by SPSS(18.0). Meanwhile, the bioinformatics analysis of GHSR gene was conducted by online software.【Result】The result showed that two SNPs (G200A and T628C) were detected in exon 2 and 3′UTR but no SNPs in exon1 and 5′UTR of GHSR gene, respectively. G200A was a sense mutation site and devided into three genotypes GG, GA and AA by PCR-SSCP. Genotype GG was the dominant genotype and allele G was dominant allele with the frequencies of 0.6731 and 0.5243 in Guizhou white goat and Guizhou black goat (♀) groups instead of allele A in Guizhou black goat (♂). All populations were moderate polymorphic in G200A site (0.25＜PIC＜0.50). G141A was found in intron 2 and two sense mutation sites (T78C and C14T) were identified in exon2 and exon4 of GHRL gene, respectively. There were two genotypes (CT and CC) in C14T site, genotype CC was the dominant genotype, which the frequency were 0.7692, 0.9417 and 0.9390, allele C was dominant allele with the frequency of 0.8846, 0.9709 and 0.9695, respectively. These populations were low polymorphic (PIC＜0.25). χ² test indicated that SNPs(G200A and C14T) fit with Hardy-Weinberg equilibrium in the population (P＞0.05). Correlation analysis revealed that G200A site was significantly associated with body weight, body height, body length and hucklebone width (P＜0.05), GG genotype was superior genotype. Additionally, body weight, body height and chest girth of individuals of Guizhou black goat (♂)with genotype CC was significantly higher than those with genotype CT(P＜0.05) in C14T site. Meanwhile, the combined genotype was significantly associated with body height and chest girth, the individuals with CCGG genotype had significantly better body height than that of genotype CCAA (P＜0.05). Bioinformatics analysis indicated that no core promoter regions and CpG island were found but a CAAT-box and several important transcription factors in 5′UTR region of GHSR gene. G200A had led to the change of secondary structure of mRNA. The secondary structure of protein was mainly α-helix (48.90%). Tertiary structure and transmembrane helix prediction showed that GHSR protein was a membrane protein.【Conclusion】The results revealed that SNPs and combined genotype of GHSR and GHRL gene had effect on growth traits in goat, G200A and C14T sites could be used as effective genetic markers for goat molecular breeding.

【Objective】 The objectives of this study were to investigate the effects of nano-zinc supplementation in dietary on the semen quality parameters, the activities of antioxidant enzyme in seminal plasma and the expression of copper zinc superoxide dismutase in epididymis of ram lambs, and to analyze the correlation between semen quality parameters and the activities of antioxidant enzyme.【Method】Sixteen 9-month-old Jinzhong ram lambs with good health and approximate weight were randomly divided into 4 groups, fed with a basal diet with supplementation of 0, 50, 100 and 150 mg·kg^-1 DM nano-zinc, respectively. The experimental period was 90 d. Semen was collected on 78 d and 79 d in consecutive two days, samples of 100 µL fresh semen was used to analyze semen quality parameters, the rest of fresh semen sample was centrifuged at 2 000 r/min for 10 min, the supernatant (seminal plasma) was collected for measurement of the activities of copper zinc superoxide dismutase, glutathione peroxidase and total anti-oxidation competence. All rams were castrated for collection of epididymis caput, corpus and cauda at the end of experiment. Expression of Cu-ZnSOD protein in epididymis was detected and located by immunohistochemistry. The mean optical density was analyzed with Image Pro Plus 7.0 software.【Result】The results showed that there was no significant difference in ejaculate volume between the treatment groups and control group (P＞0.05). Compared with the control group, the groups with supplementation of 50 mg·kg^-1 or 100 mg·kg^-1 nano-zinc had higher sperm concentration, sperm motility and membrane integrity, and less percentage of abnormal sperm. The group with supplementation of 150 mg·kg^-1 had higher membrane integrity than control group, while it had no significant difference in sperm concentration, sperm motility with control group. The rams in groups with supplementation of 50 mg·kg^-1 or 100 mg·kg^-1 nano-zinc also had higher activities of Cu-ZnSOD, GPxs and total anti-oxidation competence, and less MDA concentration in seminal plasma. The rams in group with supplementation of 150 mg·kg^-1 had higher total anti-oxidation competence than the control group, while they had no significant difference in activities of Cu-ZnSOD, GPxs and MDA content with rams in the control group. The result of immunohistochemistry showed the positive signals were detected in pseudostratified columnar epithelium in epididymis caput and corpus, and simple columnar epithelium in epididymis cauda. The order of expression of Cu-ZnSOD protein in epididymis from rams in the treatment group was corpus＞caput＞cauda, while those in the control group was caput＞corpus＞cauda. The mean optical density of Cu-ZnSOD was higher in the group with supplementation of 50 mg·kg^-1 or 100 mg·kg^-1 nano-zinc in epididymis caput and corpus. The activity of Cu-ZnSOD in seminal plasma had a positive correlation with sperm concentration, sperm motility and membrane integrity, and a negative correlation with percentage of abnormal sperm.【Conclusion】The optimal supplementation of 50 mg·kg^-1 or 100 mg·kg^-1 nano-zinc in diet could improve semen quality and seminal plasma anti-oxidase activities, and the expression of Cu-ZnSOD in epididymis of rams. The activity of Cu-ZnSOD in seminal plasma could be regarded as index of evaluation of semen quality in sheep production. The further researchs need to be done in molecular mechanism of trace element zinc regulation on semen quality.

【Objective】 The objective of the present study is to identify the target genes of miR-663 and investigate the role of miR-663 in melanin synthesis in alpaca melanocytes.【Method】 The potential targets and binding sites of TGF-β1 were predicted and analyzed by Targetscan, RNAhybrid and RNA22. The similarity of 3′UTR of TGF-β1 sequences from various species was analyzed by DNAMAN. The dual-luciferase construct of pmirGLO-TGF-β1-3′UTR was created by inserting partial TGF-β1 3′UTR into the pmirGLO vector by SacⅠ and XbaⅠ restriction sites. The regulation of TGF-β1 by miR-663 was validated by co-transfecting pmirGLO-TGF-β1-3′UTR construct with miR-663 mimic into 293T cells. The over-expression of miR-663 was achieved by transfecting melanocytes with miR-663 mimic. The mRNA and protein levels of TGF-β1, Smad3, Smad4, Smad7 and β-catenin in melanocytes transfected with miR-663 mimic were analyzed by qRT-PCR or Western blotting, respectively. The effects of miR-663 on melanin synthesis were evaluated by measuring the melanin content of the cells.【Result】There are 68 potential targets for miR-663 predicted by bioinformatics, including 74 conserved binding sites and 44 less conserved binding sites. DNAMAN analysis showed that all 3′UTR sequences of TGF-β1 from analyzed species are highly conserved and enriched potential target sites. One of the potential targets of miR-663 is TGF-β1, which is involved in the development of hair follicle as well as melanin pigmentation. The alpaca 3′UTR sequence of TGF-β1 contains three miR-663 potential binding sites. To confirm the regulation of TGF-β1 by miR-663 through its 3′UTR, a dual-luciferase reporter vector pmirGLO-TGF-β1-3′UTR was successfully constructed and co-transfected into 293T cells with miR-663 mimic. The luciferase assay experiments showed that the luciferase activity was 31.01% lower in cells co-transfected with pmirGLO-TGF-β1-3′UTR and miR-663 mimic than that in control cells, suggesting TGF-β1 is a direct target of miR-663. When miR-663 mimic was transfected into melanocytes, the mRNA level of miR-663 increased to 345% but those of TGF-β1, β-catenin and Smad4 were reduced by 89%, 41%, and 34%, respectively. The protein level of TGF-β1 was reduced by 21%. The contents of melanin were significantly reduced by 42%.【Conclusion】TGF-β1 is a direct target gene of miR-663. Overexpression of miR-663 led to the decreased expression of TGF-β1 both at protein and mRNA levels. The miR-663 may influence/affect synthesis through regulation of TGF-β1 directly as well as TGF-β/Smad and Wnt signal pathways indirectly in skin melanocytes of Alpaca.

【Objective】 It is important to minimize the influx of cadmium (Cd) to the human food chain through consumption of agricultural products. The characteristics of the uptake and distribution of Cd in pollution-safe materials were studied to provide Cd safety rice germplasm resources. 【Method】 A field experiment was conducted to investigate the differences of 56 rice parent materials in Cd uptake and distribution in polluted paddy field. According to cluster analysis, the pollution-safe materials were chosen by the Cd content of the brown rice, and Cd uptake and distribution characteristics of rice parent materials were analyzed.【Result】There were significant differences in the Cd contents and Cd accumulations at tillering stage(CV=44.05% and CV=50.21%), booting stage(CV=23.57% and CV=28.62%) and mature stage(CV=44.98% and CV=44.69%) among the 56 rice parent materials when the field soil Cd content was 13.89 mg·kg^-1. Meanwhile, the Cd contents in brown rice ranged from 0.15 to 1.77 mg·kg^-1 among the parent materials, the ratio of maximum and minimum value reached 11.80, and the minimum value of Cd content was lower than the National Food Safety Standard. The 56 rice parent materials were divided into pollution-safe materials, general materials and high accumulation materials depending on the Cd content of brown rice. The Cd content of brown rice of pollution-safe materials was 0.2 mg·kg^-1 which was significantly lower than that of the general materials (0.65 mg·kg^-1) and the high accumulation materials (1.57 mg·kg^-1). Moreover, the lowest Cd contents of chaff and grain partition coefficient were also observed in pollution-safe materials. Shoot Cd contents in the three kinds of materials were significantly decreased with the growth stage prolonged. Furthermore, shoot Cd contents in the pollution-safe materials were significantly lower than that of the general materials and high accumulation materials at tillering, booting and mature stages. Especially, the Cd contents in shoot of the general materials and high accumulation materials were 1.35 and 3.39 times higher than the pollution-safe materials at mature stage. The pollution-safe materials exhibited significantly lower Cd accumulations in shoots compared to the general materials and high accumulation materials at the three growth stages. The maximum differences among the three kinds of materials were observed at maturity stage. The Cd accumulations in shoots of the general materials and high accumulation of materials were 2.23 and 3.86 times higher than that of the pollution-safe materials at mature stage. The maximum differences among the three kinds of materials were also observed at maturity stage. The greatest Cd accumulation in shoots of pollution-safe materials was observed at sowing-tillering stage. However, there were no difference among the three growth stages in the general materials and high accumulation materials. Due to the lower metastatic ability of Cd to grain, the pollution-safe materials have lower Cd content in grain. Meanwhile, the distribution ratio of Cd accumulations in grain was 8.11% of the total Cd accumulations in aboveground of the pollution-safe materials, which was lower than that of the general materials (11.60%) and high accumulation materials (17.59%).【Conclusion】Among the pollution-safe materials, the Cd contents in the brown rice of D62B, IRBN95-90 and GRlu 17/ai TTP//lu 17_2 were lower than the National Food Safety Standard (0.2 mg·kg^-1). Thus, D62B, IRBN95-90 and GRlu 17/ai TTP//lu 17_2 can be considered as Cd safety rice germplasm resources for Cd-polluted farmlands.

【Objective】The aim of this study was to clone the full-length cDNA of sugarcane NADP⁺- dependent lsocitrate dehydrogenase, a key enzyme gene relating to tricarboxylic acid cycle (TCA) in sugarcane, to provide a candidate gene for breeding of sugarcane disease resistance and other stress tolerance.【Method】Two-dimensional electrophoresis (2-DE) was used to study the variation expression of proteins, and the differential proteins were identified by MALDI TOF/TOF. The SoNADP-IDH gene cDNA sequence was cloned from sugarcane variety GT11 using RT-PCR techniques. Online software was used to analyze the putative amino acid sequence, and qRT-PCR method was used to study the expression of SoNADP-IDH gene in different tissues and under different stresses. 【Result】 The full-length cDNA of SoNADP-IDH (GenBank accession number KF808326) in sugarcane was cloned. The full-length cDNA was 1 497 bp with an intact open reading frame of 1 239 bp, encoding a polypeptide of 412 amino acids. The bioinformatics analysis showed that the protein was a hydrophilic protein, did not contain a signal peptide, but contained four transmembrane regions. This protein was a soluble and stable protein. Secondary structure analysis showed that the protein contains alpha helix, bend region, extended strand, random coil, and alpha helix and random curl occupied most of the protein secondary structure. Online software analysis showed that the gene contains 19 phosphorylation sites, 4 N-glycosylation sites, 4 casein kinase phosphorylation sites Ⅱ, 4 N-myristoylation site of action, 6 protein kinase C phosphorylation sites and 1 tyrosine kinase phosphorylation site. Homology analysis showed that the deduced SoNADP-IDH protein was highly homologous to other IDH proteins from different species. The qRT-PCR analyses showed that the SoNADP-IDH expressed in root, stalk and leaf. Furthermore, SoNADP-IDHtranscription level was impacted under the treatment of the pathogen of ratoon stunting disease, low temperature, PEG, NaCl and ABA stresses, but the expression patterns were different. In PEG simulated drought treatment, the transcript of SoNADP-IDH showed first increase and then decrease, reached the peak at 6 h, then continued to decline, and reached the minimum at 48 h. In the 4℃ cold stress treatment, the expression of SoNADP-IDH reached the minimum at 3 h, with the extension after treatment, the expression of SoNADP-IDH increased, reached the peak at 24 h, then slightly decline at 48 h. After ABA application, the transcript of SoNADP-IDH showed a “suppression-increase-suppression-increase” pattern, the transcript of SoNADP-IDH levels decreased at 3 h, reached the peak at 6 h, to a minimum at 24 h, and rose again at 48 h. In salt-treatment, the expression of SoNADP-IDH showed an “increase-suppression-increase” pattern, reached the peak at 6 h, then began to decline sharply, down to the lowest at 24 h, then began to rise, and basically flatted with the control at 48 h 【Conclusion】 The gene SoNADP-IDH was firstly isolated and characterized from sugarcane, and the pathogen of ratoon stuning disease, low temperature, PEG, NaCl and ABA stresses impacted the expression of the gene, indicating that the SoNADP-IDH gene may play an important role in resistance to oxidative stress of sugarcane.

【Objective】 This paper aims to isolate the cDNA of GA2ox from Diospyros kaki Linn. cv. Nantongxiaofangshi, and do some preliminary study on their functions and expression level, in order to lay a foundation for further exploration of the dwarf mechanism and breeding of new dwarf cultivars. 【Method】 Total RNA was extracted from leaves of ‘Nantongxiaofangshi’ persimmon by improved CTAB method. Twofragments were identified from cDNA of ‘Nantongxiaofangshi’ by degenerate primers, and their full length cDNA were acquired by RACE amplification and named as DkGA2ox1 and DkGA2ox2, respectively. Gene structure characteristics were analyzed using the bioinformatics software. Quantitative real-time PCR (qRT-PCR) was performed to determine the expression pattern during pre-budding period, budding period, leaf expanding period, tip buds dying period, flowering period, physiological fruit-falling period, fruit coloring period, fructescence and abscission period. 【Result】 The full length cDNA of DkGA2ox1 and DkGA2ox2 were 1 318 bp and 1 267 bp, respectively, containing 5′ untranslated region (UTR) with lengths of 198 bp and 61 bp, 3′ UTR with lengths of 97 bp and 172 bp, and coding region with lengths of 999 bp and 1 005 bp which encoded 332 and 334 amino acids. The two amino acid sequences shared 73%-77% in homology compared with Populus tomentosa (JX102472.1), oleander (AY594292.1), tobacco (AB125232.1), Petunia (GU059939.1), apple (FJ571521.1), pears (JF441168.1) and grapes (JQ608472.1). The conserved structural domain analysis revealed that DkGA2ox1 and DkGA2ox2 had the typical functional domains of GA2ox protein, containing Fe²⁺ binding sites (DkGA2ox1: His-205, Asp-207, His-262; DkGA2ox2: His-204, Asp-206, His-261) and 2-oxoglutarate binding sites (DkGA2ox1: Arg-272, Ser-274; DkGA2ox2: Arg-271, Ser-273), as well as the 2OG-Fe(II)-Oxy protein domains. The protein molecular weights were 36 596.1 Da and 37 544.2 Da, respectively. Both of them are stable proteins, have no signal peptide, transmembrane domains, and significant hydrophobic region, as well as belong to C19-GAoxs. After construction of transient expression vector and onion epidermal cell transformation, subcellular localization assays showed that the GA2ox1 protein was located in the nucleus and cytoplasm. The quantitative RT-PCR results showed that the highest expression levels of DkGA2ox1 and DkGA2ox2 were detected in florescence, and all higher than those in vigorous cultivar ‘Dafangshi’ during all the 7 phenological periods. 【Conclusion】 The expression of gibberellin 2-oxidase genes in ‘Nantongxiaofangshi’ is related with the dwarf trait.

【Objective】A rice mutant, dwarf and deformed flower 2 (ddf2) , was characterized in the present paper. These results will provide a foundation for map-based cloning and functional analysis of the DDF2 gene. 【Method】 The ddf2 mutant was derived from tissue culture mutation generations of indica restorer 602. At the heading stage, the plant height, panicle length, width and length of leaves and internodes of the ddf2 mutant and wild type were measured. Moreover, the phenotype and histology analyses of stem, leaf and spikelet were conducted. A population containing 1 024 F₂ mutants from the zhonghua11/ddf2 cross was used to map the DDF2 gene. The expression of candidate genes were detected by real time PCR. 【Result】 The leaves and internodes of ddf2 mutant were all much shorter and thinner than those of the wild type, and the panicleof ddf2 mutant was shorter than that of the wild type. According to the histology analysis, the width between two large veins of leaf was decreased dramatically in ddf2 mutant, while no obvious difference in the numbers of large and small veins was observed between the ddf2 mutant and the wild type. Further analysis showed that the size and number of mesophyll cells in ddf2 leaves were all reduced significantly. Furthermore, parenchyma cells were significantly reduced in size and number in ddf2 stems, while there was no difference in the number of vascular bundles in stem between the ddf2 mutant and the wild type. These results indicated both the cell division and cell expansion were inhibited in ddf2 leaves and stems. Additionally, the spikelet and floral organs in ddf2 mutant display serious defects. In ddf2 florets, the lemmas were distorted while the paleas were degraded in whorl 1. The stamens were degenerated seriously or transformed into pistil-like organs in whorl 3. Elongated sterile lemma and extra florets were often observed in some ddf2 spikelets. Genetic analysis indicated that ddf2 was a nuclear recessive gene. By using 1 024 F₂ mutants from the zhonghua11/ddf2 cross, the DDF2 gene was mapped between In/Del markers S-11 and S-14 on the short arm of chromosome 11 near the centromere, with genetic distance of 0.049 cM and 0.098 cM, respectively, and an approximate physical distance of 90.295 kb. The gene was co-segregating with another In/Del marker S-24. There were 12 MSU annotation genes within the mapping region. Based on the qRT-RCR, it was found that the expression of LOC_Os11g17600, encoding an exocyst complex component Sec3_C protein, was down regulated in the ddf2 mutant. Then, the LOC_Os11g17600 was considered as candidate gene of DDF2.【Conclusion】DDF2 is considered to be a novel gene controlling both stem, leaf and floral organ development.

【Objective】 CORONATINE INSENSITIVE 1 (COI1) is a critical component of jasmonate (JA) receptor complex. The objective of this study is to investigate the spatial expression pattern and regulatory roles of COI1 gene in oilseed rape(Brassica napus), a staple oil crop in the world. 【Method】 The COI1 genes in oilseed rape(B. napus) and its parental species B. rapa and B. oleracea were analyzed based on the genomic data. The spatial transcription pattern of COI1 in oilseed rape was analyzed by RT-PCR with specific primers according to the conserved region of COI1 gene homologs. Then, a cDNA fragment of the conserved region was cloned into vector pTRV2 of the tobacco rattle virus-based VIGS (virus-induced gene silencing) system. And, VIGS techniques were applied to silence COI1 genes in oilseed rape. The rape plants shown to have COI1 genes silenced were used to investigate male fertility and aphid resistance. 【Result】 Analysis of the genomic data of B. rapa, B. oleracea, and B. napus revealed that the genomes of B. rapa and B. oleracea contained 7 highly homologous COI1 genes, which could be classified into 4 subgroups, i.e., COI1a, COI1b.1, COI1b.2, and COI1c, while 8 COI1 genes were presented in the genome of B. napus. Transcriptional assay revealed that the expression of COI1 in oilseed rape is tissue specific. A 505bp fragment of the conserved region of COI1 was introduced into the VIGS vector pTRV2 to develop COI1-silenced plants via VIGS techniques. Twenty-five plants with the transcripts of COI1 down-regulated by over 70% were identified by transcriptional analyses, from which the ten plants with the lowest expression level of COI1 were selected to investigate male fertility and aphid resistance. The results showed that the fertility of COI1-silenced plants was extremely impaired, with no seeds in their siliques. Furthermore, the filaments of COI1-silenced plants were shorter than those of control plants, and over 80% pollens of the COI1-silenced plants were irregular in shape. Aphid resistance assay showed that aphids grew and reproduced much faster on the COI1-silenced plants than on the control plants cultured under the same condition. 【Conclusion】 This study revealed that the expression of COI1 gene in oilseed rape is tissue specific. VIGS-induced silencing of COI1 greatly impaired the male fertility and decreased the aphid resistance of oilseed rape. This work has provided an important start point to dissect the mechanism of JA signaling in oilseed rape.

【Objective】The aim of this study was to illuminate the sensibility of the exogenous hormones to plant height of a dwarf germplasm Aidianyehuangma so as to determine which kind of hormone sensitivity this germplasm belongs to, which will facilitate dissecting the relationship between the genetic basis of plant height and hormonal regulation in jute. 【Method】 Firstly, the performance of important agricultural traits, e.g. plant height, were compared between Aidianyehuangma and the variety Huangma 179 with normal plant height. Secondly, the dwarf germplasm of Aidianyehuangma was treated with different concentrations of exogenous GA₃, IAA and BR. And the hypocotyl length in the Ms culture media and plant height variation in the field were investigated. Meanwhile, the hypocotyl length of Huangma 179 and Aidianyehuangma on the GA₃ culture media was evaluated. Thirdly, the effects of continuous spraying of 100 mg·L^-1 GA₃ every 3 days on plant height of Aidianyehuangma were analyzed. Furthermore, the activities of α - amylase of Aidianyehuangma were analyzed after it was once treated with GA₃. 【Result】 The comparison analysis of important agricultural traits showed that the plant height of Aidianyehuangma (1.89 m) was significantly lower than that of Huangma 179 (4.28 m) without exogenous hormone treatment, indicating that Aidianyehuangma is a typical dwarf germplasm in jute. After being treated with MS culture media of 0.1 mg^.L^-1 exogenous IAA and BR, the hypocotyl length of Aidianyehuangma did not show a significant difference with that of the control. However, after being treated with MS culture media of 0.1 mg·L^-1 exogenous GA₃, the hypocotyl length of Aidianyehuangma showed a significant difference compared with Aidianyehuangma without being treated with GA₃, and the hypocotyl length of Aidianyehuangma and Huangma 179 was similar to that of Huangma 179 without being treated with GA₃. It suggested that the plant height of Aidianyehuangma is sensitive to exogenous GA₃ while that of Huangma 179 is not sensitive to the exogenous GA₃. After being treated with different concentrations of exogenous hormone in the field, plant height of Aidianyehuangma with high concentration (100 mg·L^-1) of GA₃ was significantly higher than that with the other two lower concentrations of GA₃. By contrast, plant height of Aidianyehuangma treated with different concentrations of exogenous IAA and BR did not show a significant difference with that of the control. It further suggested that Aidianyehuangma is a GA₃ sensitive dwarf germplasm. After being treated with 100 mg·L^-1 exogenous GA₃ continuously in the field, the plant height of Aidianyehuangma showed a significant difference in comparison of Aidianyehuangma without being treated with GA₃, reaching a significant level, and appoached that of Huangma 179. It showed that plant height of Aidianyehuangma could be resumed using exogenousGA₃. After Aidianyehuangma was once treated with GA₃, α- amylase activity of Aidianyehuangma in the first 3 days began to increase in comparison with that before treatment, and enhanced significantly in the first 6 days, which indicated that the exogenous GA₃ could increase the α- amylase activity of Aidianyehuangma. 【Conclusion】Aidianyehuangma is a typical dwarf germplasm in jute. The plant height of Aidianyehuangma is not sensitive to exogenous IAA and BR, but sensitive to the exogenous GA₃, and can be resumed to normal plant height with continuous exogenous GA₃ treatment. The exogenous GA₃ could increase the α- amylase activity of Aidianyehuangma. Aidianyehuangma is a GA₃ sensitive dwarf germplasm.

Mapping crop patterns with remote sensing is of great implications for agricultural production, food security and agricultural sustainability. In this paper, the theoretical basis behind the mapping was summarized, mapping methods were classified into several categories, characteristics and applicabilities of different mapping methods in the latest decade were discussed intensively, and some important directions and priorities for future studies were proposed. Currently, spectral, temporal and spatial features are the major theoretical bases for crop pattern mapping. The mapping method based on single imagery is characterized by its simple implementation, but with difficulty of capturing imagery at the best time for distinguishing different crops. Instead, the mapping method based on time-series of imagery can make full use of temporal features and is thus widely used for crop mapping, among which the methods using multiple features are more suitable than the ones using a single feature for regions with complicated planting structure. To some extent, feature-oriented statistical modeling method can resolve the mixed-pixel problem but its robustness needs to be improved. Furthermore, large-scale crop pattern mapping can be done by combining the remote sensing and agriculture statistics. However, due to coarse resolution, the derived maps show poor region suitability. Future crop pattern mapping should target at developing “a map of crops”, the emphasis must be put on covering more crop types, enlarging the mapping areas, utilizing the superiority of blending multi-source data, strengthening the data preprocessing, optimizing the feature extraction and classifier selection, and improving the temporal and spatial scales of crop pattern mapping so as to better meet the needs of multi-faceted agricultural applications.

【Objective】 The objective of this paper was to map the cropping systems of China using ten day NOAA- AVHRR- NDVI data of 1986 and 1996 years, and SPOT-VEGETATION-NDVI data of 2000. In order to extract the cropping systems information, the accuracies of the twice difference algorithm and Fourier transform method were compared. The performed better method was applied to map the cropping systems distributions for each year. On the basis of the cropping systems distributions, the changes of cropping systems in recent years would be known. Some meteorological data were used to reveal the possible reason of the change of cropping systems. The results were expected to provide information about the crop yield changes and their reasons.【Method】First, the cropping area was subset from the global land cover map provided by the European Space Agency (ESA GlobCover). Second, the cropping system distribution of 1986 was mapped using twice difference algorithm and Fourier transform method by ENVI and ArcGIS software. The accuracies of the two methods were compared using the cropping system information in the “Vegetation Map of China” at the scale of 1:4 000 000 as a reference. The cropping systems distributions of 1996 and 2006 were mapped by the twice difference algorithm. The distributions were compared to extract the change information of cropping systems. At last, the accumulated temperature indicators of the zero-grade zone were used to analyze the possible impact of the climate change on the cropping systems change.【Result】The twice difference algorithm and the Fourier transform both provided reasonable accuracy for mapping cropping system of China. The area of one crop a year was mainly in the northeast area, the two crops a year was distributed in Henan, Hebei, Shandong, and Anhui provinces, and the three crops a year was covered by the area south to the Hubei Province. The accuracy of twice difference algorithm (overall accuracy was 76.5% and Kappa coefficient was 0.64) was a little higher than the Fourier transform (overall accuracy was 69.4% and Kappa coefficient was 0.51) in this research. The northern limit of the two crops a year moved from Huludao, Liaoning Province in 1986 to Benxi and Shenyang, Liaoning Province in 2006. The western limit of it also moved from Pingliang, Gansu Province, Baoji, Shanxi Province in 1986 to Linxia, Ningxia Hui Autonomous Region and Haidong, Qinghai Province in 2006. The cross transformation was so complicated that the area of the one crop a year decreased by 330 thousand square kilometers, two crops a year increased by 360 thousand square kilometers, and three crops a year increased by 40 thousand square kilometers. The contour line of the accumulated temperature of the 4 200ºC and 6 100 ºC also moved to north and west in the studied years. The results may imply that the temperature is a main impact factor of changes of cropping systems.【Conclusion】Remote sensing data based on time sequence could be used to monitor real change of cropping system in China. The twice difference algorithm is a valid method to extract the crop systems information in China. The limit of the cropping system, especially for the two crops a year moved northwards and westwards. This movements resulted in the large scale decrease of one crop a year and significant increase of two crop a year, while the three crops a year did not change much. The global warming had an effect on the cropping system changes in China. Analyzing the distribution and change of cropping system in China by remote sensing data can not only help us understand and evaluate the crop production and production potential timely and accurately, but also benefit the agriculture-related departments for drawing up effective agricultural policies.

【Objective】 In the context of climate change, China needs to address critical challenges relating to the agricultural development and food security. Wheat is one of China's three major grain crops, which plays an important role in China’s food security. The variation of potential light-temperature suitable cultivation zone of winter wheat in China under climate change was studied, which would provide an important scientific reference for the rational distribution of winter wheat. 【Method】 In this research, the ‘correction step by step’ method was used to calculate the potential light-temperature yield of winter wheat in each year. The year 1981 was taken as a time node and divided the period 1961-2010 into two sub-periods. Combined with the indices of the potential light-temperature yield level and the stability, the variation of the potential light-temperature yield level and the stability zones were investigated. In summary, the variation of the boundaries and distributions of the potential light-temperature suitable cultivation zones of winter wheat in China under climate change was studied. 【Result】 Compared with the period 1961-1980, during 1981-2010, the percentage of the very high and high potential yield zones in the study area increased by 6.33% and 7.42%, respectively, but the percentage of moderately high and marginally high potential yield zones decreased by 10.50% and 3.24%, respectively. During 1981 to 2010, the percentage of the very stable potential yield zone in the study area decreased by 25.76%, but the percentage of stable, moderately stable and marginally potential yield zones increased by 12.09%, 10.34% and 3.31%, respectively. The boundary of the very suitable zones moved northward and westward. The southern boundary of the very suitable zones moved northward from the central part of Anhui Province to the northwest portion of Shandong Province, and the western boundary of the very suitable zones extended westward from the southwest part of Shanxi Province to the northwest part of Shaanxi Province; The boundary of the suitable zones moved northeastward, and the maximum spatial displacement was in the eastern part of Sichuang Province to the southwest part of Shaanxi Province, moving by about 835.63 km. The spatial displacement of the potential suitable cultivation zone of winter wheat caused the changes of the winter wheat actual cropping area. Compared with the period 1961-1980, during 1981-2010, the percentage of the very suitable and suitable zones of winter wheat in the study area decreased by 4.49% and 4.75%, respectively, while the percentage of moderately suitable and marginally suitable zones increased by 7.17% and 2.06%, respectively. The potential yield per unit area increased by about 513 kg·hm^-2 in average in each suitable cultivation zone. However, the total potential yield reduced by 1 504×10⁴ t in the study area compared with 1961-1980 because of the reduction of the area in the very suitable and suitable zones of winter wheat, which caused the total potential yield in these two cultivation zones reduced by 5 108×10⁴ t and 5 896×10⁴ t, respectively. 【Conclusion】 Under the background of climate change, the percentage of the very high and high potential light-temperature yield zones of winter wheat increased, while the very stable potential light-temperature yield zone decreased. The very suitable and suitable zones of winter wheat in the study area decreased because of the decrease of stable; the boundary of the very suitable zone moved northward and westward, and the boundary of the suitable zone moved northeastward. The potential light-temperature yield per unit area increased because of the spatial displacement of the potential light-temperature suitable cultivation zone, which was beneficial to the improvement of the winter wheat yield. However, due to the reduction of the area in the very suitable and suitable zones of winter wheat, the total potential yield of winter wheat in China reduced. In the regions where the suitability of potential light-temperature yield has changed, the wheat production should be combined with the irrigation to ensure the high and stable yield.

【Objective】Crop variety breeding was result from nature and artificial selection under climate change background. Huanghuai wheat region is the largest wheat region in China, that plays an important role for ensure wheat production and food security in China. The changes of agronomic traits of winter wheat and temperature in Huanghuai dryland in the past 30 years were studied in order to provide a theoretical basis and technological support for dryland wheat breeding to adapt to climate changes in the future. 【Method】 The agronomic traits data of control varieties and temperature data of Linfen in Shanxi province in the past 30 years were analyzed. Linfen is the national representative Huanghuai dryland winter wheat regional test site. The regularity of the main temperature traits of Linfen city, such as the average temperature and over 0℃ accumulated temperature in winter wheat growth, and agronomic traits of control varieties change trends were analyzed. The relationship between agronomic traits and meteorological elements were also analyzed by correlation, multiple regression and path analysis. 【Result】 The average temperature，over 0℃ accumulated temperature，the highest and lowest temperature was increased slowly year by year in wheat growth period. In wheat growth period, the average temperature was increased average 0.05℃ per year, over 0℃ accumulated temperature was increased average 21.9℃per year，there was positive relationship between the average temperature and over 0℃ accumulated temperature. Over 0℃ accumulated temperature change can reflect temperature change in whole wheat growth period. The highest and lowest temperature was increased 0.02℃and 0.16℃per year respectively. The average temperature and over 0 ℃ accumulated temperature was increased markedly from sowing to before the winter, the average temperature and over 0℃ accumulated temperature was increased significantly at vegetative growth stage, but were increased lightly and had a downward trend at reproductive stage. With climate warming, strong winter variety with more tillers was replaced gradually by winter and weak winter variety with moderate tiller. Agronomic traits of control variety were changed not much from 1986 to 1996, and changed greatly from 1997 to 2007, and changed markedly from 2007 to 2014, this change trend consist with temperature change trend in wheat growth period. In evolution of agronomic traits of Huanghuai dryland control varieties, the thousand-kernel-weight and kernels per spike was increased average 1.57% and 3.39% per year respectively, but the effective ear numbers and plant height as well grain yield were decreased average slowly 0.16% and 1.29% per year respectively. There was a significant positive correlation between grain yield and plant height (0.684**), effective ear number (0.531**) and thousand-seed-weight (0.541**). There were 46.73%, 26.17% and 3.26% variation of grain yield was decided by three main factors such as plant height and over 0℃ accumulated temperature as well as the average temperature from standing to jointing stage respectively. In Huanghuai dryland wheat high-yield breeding evolution, the over 0℃ accumulated temperature from standing to jointing stage and plant height had a higher positive effect on grain yield, but the average temperature from standing to jointing stage had a negative effect on grain yield. 【Conclusion】 The climate warming had a strong impact on agronomic traits evolution in Huanghuai dryland winter wheat region, selecting new wheat varieties with agronomic traits of middle plant height, middle effective ears numbers, more kernels per spike, and high grain weight is the improvement direction of middle-high yield and drought-resistant varieties in Huanghuai dryland wheat region for adapting to climate change in the future.

【Objective】The objective of this study is to clone the ORF (open reading frame) of chemosensory protein 1 (CSP1) from Bemisia tabaci biotype Q, and characterize the binding profiles of CSP1 with some candidate plants volatiles.【Method】 By means of full-length ORF primer, reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the full-length ORF BtCSP1, which was then cloned into the pET-30a (+)/BL21 (DE3) prokaryotic expression vector after double enzyme digestion. The recombinant BtCSP1 protein was expressed and purified by the method of Ni²⁺-agroase affinity chromatograph. The protein concentration was measured with Bradford method. The competitive fluorescence assay was used to analyze the binding properties of BtCSP1 with general plant volatiles with different chemical structures. As a suitable fluorescence reporter in studies of insect GOBPs’ function in vivo, N-phenyl-1-naphthylamine (1-NPN) was used to titrate the BtCSP1 solution until the fluorescence emission peak at maximum wavelength of BtCSP1 completely quenched. Then all plant volatiles were added into BtCSP1-1-NPN complex, respectively. The dissociation constants K_D data represented the affinity of BtCSP1 with ligands were calculated by the Scatchard equation. 【Result】 BtCSP1 protein was successfully expressed after induction of 1 mmol·L^-1 of IPTG, then purified by Ni²⁺ affinity column with gradient imidazole as washing solutions, finally dialyzed sufficiently using PBS buffer. The working concentration of BtCSP1 was diluted to 1.5 µmol?L^-1. In the competitive fluorescence assay, the dissociation constants K_1-NPNand the number of binding sites n of BtCSP1 and 1-NPN were 2.78 μmol·L^-1and 0.82, respectively. It indicated that the binding of BtCSP1 with 1-NPN is strong and the binding ratio is almost 1:1. In all candidate chemicals, some plant volatiles preferred to bind with BtCSP1 with high affinity, such as 3-carene, 4-cymene, HL-cis-3-hexen-1-ol and α-pinene, which are reported to be related to the repellent characteristics of B. tabaci. Their binding affinity with BtCSP1 was strong, with the dissociation constant K_D of 26.47, 39.43, 54.01 and 83.46 μmol·L^-1, respectively. Especially 3-carene reduced the relative fluorescence intensity of 1-NPN by about 40% at the concentration of 200 μmol·L^-1. 【Conclusion】 BtCSP1 exhibited a strong binding affinity with some repellent plant volatiles, indicating that BtCSP1 may be involved in the recognition of the repellent plants of host selection in the process of expanding into the invasion fields.

【Objective】Root border cells (RBCs) are released from the root cap as individual cells or a group of attached cells. The mucilage of root border cells acts in a manner similar to that of neutrophil extracellular traps (NETs) in defense, thus, known as border cell extracellular traps (BETs). The extracellular DNA (exDNA) and proteins are components of border cell mucilage, which have been considered to play a vital role in protecting root tip from biotic and abiotic stresses. Allelopathy is one of the successful mechanisms of exotic plant for invading. The objective of this study is to reveal the roles of the exDNA and extracellular proteins of root border cell mucilage in resistance to allelochemical stress from an invasive plant, Chenopodium ambrosioides L..【Method】 Experiments were performed with maize (Zea mays L.) ‘Yayu26^#’, a widely grown crop in the introduced habitats of C. ambrosioides under aeroponic culture with agar medium. The developmental characteristics of root border cells in maize were investigated. And the roots were treated with volatile oil from C. ambrosioides and its two main components, α-terpinene and cymene, respectively, then the experiments were divided into two groups. In one group, the root border cell viabilities and relative areas of root border cell mucilage were measured directly. In another group, the exDNA or extracellular proteins of root border cell mucilage were degraded by DNase I or protease, and then the cell viabilities and relative areas of root border cell mucilage of maize were studied.【Result】 The first border cell formation almost synchronized with root tip emergence in maize. The number of border cells increased with the development and elongation of the root. When the root length was about 30 mm, the number of border cells reached the maximum, that was about 6 130, then declined slightly to a stable level. Meanwhile, the activities of pectin methylesterase were correlated with root length negatively (P＜0.01). Once the emerging of root, the activities of pectin methylesterase reached the top, and then declined into a low level. The cytotoxicity from volatile oil, α-terpinene and cymene resulted in significant decrease of the viabilities of root border cells (P＜0.05). The allelopathic effects of α-terpinene and cymene were weaker than that of volatile oil. The allelopathic effects of mixed treatment group were different from the individual groups, which the viabilities of root border cells were higher than any other treatment groups, showed minimal toxicity. Volatile oil from C. ambrosioides had potential to induce the enlargement of mucilage with dose-dependent. At the dose of 5 μL, the relative areas of mucilage enlarged significantly in volatile oil treatment groups (P＜0.05). Compared with the control, α-terpinene-treated and cymene-treated induced no significant change of the relative areas of mucilage. Degradation of exDNA or extracellular proteins with DNase I or protease resulted in loss of root border cells resistance to allelochemical stress, and the viability of root border cells decreased obviously, there was less than 10% viabilities of root border cells remained in volatile oil treatment groups, while there was more than 80% in α-terpinene-treated, cymene-treated and the combined treatment of α-terpinene and cymene. 【Conclusion】Volatile oil from C. ambrosioides and its two main components, α-terpinene and cymene were cytotoxical, both of them inhibited viabilities of root border cells, while exDNA and extracelluar proteins of root border cell mucilage play an important role in resistance to allelochemical stress from C. ambrosioides.

【Objective】Ryanodine receptor (RyR), the target of diamide insecticides, is the largest known iron channel protein. The receptor is the key to the stabilization of Ca²⁺ by regulating the release of Ca²⁺ in cell. The objectives of this study are to isolate the cDNA of ryanodine receptor from the peach fruit moth Carponsina sasakii (CsRyR), and to analyze the expression of RyR mRNA at different developmental stages of this pest. 【Method】 According to the sequences of RyR gene of other species obtained from the NCBI database, degenerate and specific primers were designed for reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends PCR to clone the full-length sequences of CsRyR. The open reading frame (ORF), animo acid residues, the conserved structure domains, phylogenetic tree and other characteristics were analyzed using the bio-software. And the relative expression levels of RyR mRNA at different developmental stages (egg, larvae, pupe and adult) were investigated using quantitative real-time PCR (RT-qPCR), GAPDH was used as the reference. 【Result】 The full-length cDNA sequence of CsRyR was isolated from the peach fruit moth using RT-PCR and RACE-PCR. The CsRyR mRNA has an open reading frame (ORF) of 15 405 bp nucleotides and encodes 5 134 amino acid residues. CsRyR displays 45%-47% identity with vertebrate RyR isoforms, and 46% identity with RyR from Caenorhabditis elegans (CeRyR). And CsRyR shares 91%-94% identity with that of Lepidoptera insects and 79% identity with those of Homoptera and Diptera insects. Meanwhile, the phylogenetic tree analysis showed that CsRyR has the closest genetic relationship with insects of Noctuidae and Pyralididae.CsRyR shares common structural features with known RyRs. Six transmembrane domains (AA 4 467-5 029) are at the COOH-terminal. The sequence motif, GVRAGGGIGD, which constitutes part of the pore-forming segments of the Ca²⁺ release channels. The sites of RyR binding with Ca²⁺ and ATP (EF-hand and GXGXXG (3 motifes)) exist. Furthermore, three putative sites potentially binding with diamides were AA 183-290 (BmRyR), AA 4 610-4 655 (DmRyR) and 4 946 G (PxRyR) showed no differences in the CsRyR. And the relative expression abundance of RyR mRNA from the pupae of C. sasakii was the highest, which were 25.19, 7.73, 6.48, 4.74 and 3.58 fold compared to 1-day-old eggs, 6-day-old eggs, infant larvae, mature larvae and adults, respectively. 【Conclusion】 The full length of RyR cDNA was cloned from C. sasakii, and the mRNA expression has significant differences at different developmental stages.

【Objective】The aim of this study on the steppe soil in Qinghai Lake area is to explore the soil moisture migration and its enrichment characteristics, soil moisture distribution model, hydrological cycle and balance and reveal the characteristics of soil reservoirs, dried soil layer and its recovery conditions, and then provide a scientific basis for protecting soil water resources and steppe vegetation, construction of soil reservoir and sustainable development of grassland ecological environment.【Method】A series of experiments were performed to determine soil moisture, soil suction, infiltration rate and particle size. More than 600 soil moisture samples were collected by Light-duty Human Drill in 2009-2012. Drying weighing method was used to determine soil water content. Bicyclic infiltration method was used to determine soil infiltration rate in situ. Soil particle size was analyzed by Laser Particle Analyzer. Soil suction was measured by Tensiometers in situ.【Result】The distribution of soil water at soil profile has a certain stability, that is, in dry season or rainy season, the soil water in this area is enriched about 65% at 0-0.4 m depth and is quite shortage under 0.6 m depth. The soil suction in this area ranging from 0.17 MPa to 0.42 MPa suggests that field capacity of this area is about 20%. The infiltration rate of soil in this area varying from 1.3 mm·min^-1 to 3.0 mm·min^-1 shows that it is high and advantageous to precipitation into soil. The soil moisture is about 23% and 6.5% at 0-0.4 m depth and under 0.6 m depth, respectively. Field capacity of this area is about 20%, meaning that there is about 3% gravity water at 0-0.4 m depth of soil. The soil usually develops a dried soil layer with different levels under about 0.6 m in this area, and the greater soil thickness, the more seriously dried soil layer develops. Under 0.4 m depth of soil, the relationships between soil moisture and depth can be described by power function. The parameters of power functions (i.e. a, k) are able to well reflect the average level of soil moisture (a) and the degree of soil moisture decreased with the increase of depth (k). As the precipitation increased by about 50 mm from 2009 to 2011, the increment of soil moisture can be clearly reflected by the incremental curve of analogue function. The increment of soil moisture within the depth from 0.4 m to 0.8 m gradually decreased from 5% to 3% and was less than 3% under 0.8 m depth. The soil property is excellent in this area, but the soil moisture under 0.6m depth is close to or below 5% unavailable water of silty soil.【Conclusion】The retention and enrichment of soil water at 0-0.4 m depth is rare and this characteristic mainly depend on the low temperatures and long-term freezing of soil in Qinghai Lake area. The regulation function of soil reservoir is weak in this area due to the average thickness of soil is less than 1.5 m. To some extent, the retention and enrichment of soil water not only can enhance the regulation function of soil reservoir for vegetation in shallow root, but also can play an important role in inhibiting the occurrence of grassland desertification. The soil water balance in this area showed a weakly positive balance because of 400-420 mm annual precipitation from 2009 to 2011. Under this condition, the dried soil layer in thin soil was able to disappear completely, but could not in thick soil. The dried soil layer was recovered slowly in this area and its soil moisture recovered was less than 5.0%. Development of dried soil layer and its distribution with small depth not only show that the ecosystem in this area is more vulnerable, but also indicate this area is not suitable for growing arbor vegetation because it will consume more water.

【Objective】The objective of this experiment is to study the effects of different water and nitrogen levels on root growth and a fruit weight of greenhouse muskmelon, to explore the relationships between root growth, single fruit weight and water and nitrogen supply, and to provide a scientific basis for water and reasonable nitrogen application of local greenhouse muskmelon.【Method】According to the environment parameters (light, temperature, humidity, etc.) inside the solar greenhouse, ‘Yi pin tian xia 208’ of muskmelon was chosen as the test cultivar, irrigation application amount was determined by using modified Penman- Monteith equation. A total of 9 treatments were designed. Muskmelon plants were subjected to three irrigation treatments, that wass 70%, 100% and 130% of crop ET_c (Evapotranspiration) and three N levels, including low nitrogen level N₁ (70 kg N·hm^-2), traditional nitrogen level N₂ (130 kg N·hm^-2) and high nitrogen level N₃ (180 kg N·hm^-2). Treatments were completely arranged randomly. A greenhouse muskmelon crop was grown under mulched drip irrigation conditions to investigate the effects of different water and nitrogen levels on root growth, distribution and single fruit weight.【Result】 Most of the total root lengths were concentrated on the 0-30 cm soil layer. Total root length increased slowly as soil layer increased. Total root length, single fruit weight and water use efficiency of muskmelon descended with the rising of irrigation water amount at the same nitrogen level. The highest values of total root length and single fruit weight were observed in W₂N₂, which were 6 625.48 cm and 818.94 g, respectively. Total root length and single fruit weight of muskmelon showed a trend of decreasing after the first increasing with the rising of irrigation water amount at the same nitrogen level. Water use efficiency descended with the rise of irrigation application rate, PFPN descended with the rise of nitrogen application rate. There was a significant relationship between root length with less than 2 mm, root dry weight and yield，and the correlation reached a significant level, the more root growth, the higher yield would increase, reasonable amount of irrigation water and nitrogen fertilizer could promote root system to absorb water and nutrients, thus increasing production.【Conclusion】The change rule of total root length in the depth of root vertical direction could be simulated by the equation y=A(1-B^x), the equation determination coefficient R² reached more than 0.9. The principle component analysis method was used to evaluate the root growth of muskmelon, and the results showed that comprehensive principle component could represent 97.27% total root information, the highest comprehensive evaluation was obtained in W₂N₂. Unreasonable irrigation and nitrogen applications could result in reducing single fruit weight, characteristic parameters of muskmelon root, water use efficiency and partial factor productivity from applied N. It was concluded that the irrigation level W₂ and nitrogen level N₂ could be recommended as the best combination of water and nitrogen, which promoted the root growth, improved single fruit weight, water and nitrogen use efficiency of muskmelon in the muskmelon production under drip irrigation with plastic film mulching in the experimental area.

【Objective】 The objective of the paper is to study the expression pattern of eight genes in different nodes and the phenological period of buds development, and explore the mechanism of the time and speed of buds development on different nodes of grape branches.【Method】 In this study, the expression of eight floral genes (VvFT, VvSOC1, VvAP1, VvAP2, VvAP3, VvFUL, VvAG and VvFLC) in the buds at different branch nodes of 8-year-old ‘Fujiminori’ grapevine was examined by real-time quantitative PCR, and the phenological period of flower bud differentiation was observed. 【Result】The floral differentiation of ‘Fujiminori’ grapevine showed a remarkable characteristics of ‘beyond section’. The first bud began to form and develop from late April or early May, and the formation time of the last bud at the top was in late September. The development time of the lower nodes was five months longer than the upper nodes, but the flowering time of them was almost at the same in the second season. The buds in the upper position differentiated from the bottom upwards to the top gradually, and the process of bud differentiation at the top was shorter than that at the bottom. The expression of floral genes in buds at different branch nodes was different. The expression level of VvFT was low and showed no significant difference during the growing season. VvSOC1 , VvAP2, VvAP3 and VvFUL. The highest expression level appeared in the early period of bud differentiation, and then decreased over time. Buds on middle nodes undergo a low speed differentiation process for a long time, and the expression of floral genes kept a high level during the differentiation period. By contrast, the buds on upper and lower nodes differentiated at a higher speed, and the expression of floral genes was lower. In the end, the development of the buds on different nodes reached the similar level. 【Conclusion】 The expression levels are different on different nodes, and high expression periods were also different on different nodes. Although the peak period of floral development of lower and upper buds were relatively late, continuous development was observed in these nodes, which led to the floral differentiation simultaneously on the lower, middle and upper nodes. This may be one of the factors that lead to difference of differentiation quality between buds on different nodes.gene in the buds at different branch nodes expressed highly during all the growing season, and showed a similar expression trend. The expression of VvAP1, VvAP2, VvAP3, and VvFUL genes in the buds at lower and upper nodes showed no remarkable change, however on the middle nodes, the expression of these genes changed greatly, and appeared an obvious upgrade firstly then descending latter tendency, what’s more, in the buds on 8, 11 and 15th nodes, the expression of these genes was higher than that at other nodes. VvAG showed different expression patterns with VvAP1

【Objective】 By comparison analysis of the results of different sampling strategies and genetic distances, the method for constructing core collection of Xinjiang wild apricot based on molecular markers data was studied in order to establish the optimum core collection resources which will be beneficial to the protection and use of resources. 【Method】 Taking 135 wild apricots as materials that were come from Huocheng Daxigou, Xinyuan Boersai and Gongliu Yiligedai in Xinjiang, using UPGMA stepwise clustering method according to SM, Jaccard and Nei&Li genetic distances for initial collection, until any one sampling location was no collection into the core collection, then an allele preferred sampling strategy was used to construct wild apricot core collection and compared with the random sampling strategy. The number of lost allele and t-test of number of polymorphic loci, percentage of polymorphic loci, observed number of alleles, effective number of alleles, Nei’s genetic diversity and Shannon information index for gene diversity were used to determine the optimal building methods. T-tests of core collection, initial collection and reserve collection were conducted, and the genetic diversity of them was compared to evaluate the representative of core collections. The principal coordinate analysis method and the phenotypic traits of initial and core collections were used to confirm the core germplasm.【Result】Compared with the random sampling strategy, allele preferred sampling strategy could construct more representative core collections that with higher values of genetic diversity indexes and little polymorphic loci. According to Nei&Li genetic distance, the constructed core collection of Xinjiang wild apricot was better than by SM and Jaccard genetic distance for these core collections had high values of genetic diversity indexes. The analysis of principal coordinate analysis and phenotypic traits showed that the core collection of Xinjiang wild apricot constructed by allele preferred sampling strategy and Nei&Li genetic distance could more comprehensively represent at the genetic diversity of wild apricot initial collection. The 31 core collection resources of wild apricot includes 22.96% germplasm samples of the initial collection, the retention ratio of number of polymorphic loci, percentage of polymorphic loci, observed number of alleles, effective number of alleles, Nei's genetic diversity and Shannon information index were 92.69%, 98.83%, 99.42%, 103.26%, 109.24% and 108.31%, respectively. 【Conclusion】 The method of allele preferred sampling strategy and Nei&Li genetic distance by stepwise clustering is a suitable method for constructing Xinjiang wild apricot core collection. These results demonstrated that the 31 core collection could stand for original collection excellently, at the same time this research method of the construction of core collection has important reference values for other crops.

【Objective】 Fresh walnut fruit easily appears decay and browning, its kernel easily becomes mildewed and dehydrated during storage. The supply period of fresh walnut fruit is short and walnut used in production is often conserved by dry. The objective of this study is to provide a theoretical basis and technical method for prolonging the supply period of fresh walnut through studying its postharvest physiology and storage technology. 【Method】 ‘Xiang ling’ walnut variety was used as raw material in this study, air flow method was used to analyze the respiratory intensity of fresh walnut with husk and fresh walnut without husk. Moreover, the respiration type was also studied. Before storage under the condition of temperature (1.0±0.5)℃, RH 90%-95%, the materials were subjected to storage in vacuum-package with plastic bag, modified atmosphere package with plastic bag, controlled atmosphere storage of text box, respectively. A contrast was prepared without any treatment. To study the effect of gas composition on postharvest physiology and storage technology of fresh walnut fruit, moisture content, reducing sugar content, lipid content, protein content, MDA content and oil acid value, iodine value, peroxide value of fresh walnut kernel were measured during storage. Related enzymes of fresh walnut kernel such as LOX activity, SOD activity and CAT activity were determined, too. Moreover, green husk browning index of fresh walnut fruit was evaluated in different periods of storage. After storage for 120 d, walnut shell color, endotesta color, kernel flavour of total fresh walnut fruit were evaluated, respectively. Then, the appearance of fresh walnut husk, shell and kernel were taken photo. 【Result】 The results showed that the fresh walnut fruit with green husk is respiration climacteric fruit and fresh walnut without green husk is non-respiration climacteric fruit. The content of moisture, lipid, and oil iodine value presented a reduced trend, but the acid value, peroxide value, LOX activity, and MDA content of fresh walnut kernel showed a increased trend and SOD activity, CAT activity, reducing sugar content, protein content showed no visible change. The contrast treatment showed the worst preservation effect and green husk browning index reached 0.27 after storage for 40 d. However, the green husk browning index of vacuum-package with plastic bag storage was only 0.18 after storage for 120 d. The result also showed that the treatment of vacuum-package with plastic bag could effectively prevent browning of green husk, maintain the moisture of kernel, slow down the change rate of oil acid value, iodine value and peroxide value, keep the activities of SOD and CAT at a high level, inhibit the generation of MDA. By evaluating the sensory quality after storage for 120 d, the treatment of vacuum-package with plastic bag gained the highest score in every single project. 【Conclusion】 The lower the concentration of O₂ in storage circumstances, the better the effect of conservation. After plucked with carpopodium in walnut commercial mature period, pre-cooled at 4.0℃ for 3 d, and vacuumed with LDPE plastic bag using vacuum pack machine, the storage life of walnut could reach 120 d at（1.0±0.5)℃ and RH 90%-95%. Its pericarp browning index of fresh walnut was low, and the color, aroma, taste and shape remained good.

【Objective】This study was conducted to get a comprehensive understanding of miRNAs expression in sheep ovary, and to analyze their expression differences between sheep producing single lamb and twins, thus providing a basis for exploring the role of miRNAs in the regulation of fertility. 【Method】 Multi-species miRNA microarray was used to profile ovarian miRNAs expression in sheep. Firstly, single-lamb-producing and twin-producing ewes were selected. Ovaries were collected after estrus for total RNA extraction, and small RNA fragments were isolated and hybridized with miRNA microarray. Then data analysis was performed to obtain sheep ovarian miRNAs profile. Differentially expressed microRNAs were selected by bioinformatics software with the standard of q-value ≤ 5% and Fold Change ≥2 or ≤0.5, and q-PCR technique was employed to testify the microarray results. Two different methods, microT and miRDB, were used, respectively, and then data were merged and to predict target genes. Finally, online software was used to perform GO function annotation for target genes. 【Result】 Among all the detected miRNAs, a total of 5448 miRNAs were detected in sheep ovaries, of which 22 were specifically expressed in single-lamb-producing group, while 15 were in twin-producing group; For the miRNAs reported in sheep, expression differences were compared between two groups, and obtained 11 differentially expressed miRNAs, with 7 down-regulated and 4 up-regulated. Tow miRNAs were randomly selected to perform q-PCR validation, and the results were consistent with the microarray results, illustrating that the microarray results were accurate and credible. Target genes were obtained for 7 miRNAs: oar-miR-370-5p, oar-miR-376b-5p, oar-miR-381-5p, oar-miR-412-5p, oar-miR-541-3p, oar-miR-544-5p and oar-miR-1185-5p，and the number of target genes for each miRNAs was: 115, 71, 1, 5, 8, 135 and 23. The GO annotation results showed that target genes of miR-376b-5p and miR-1185-5p are mainly involved in forming intracellular components and organelles; in the molecular function category, the vast majorities of genes are the connection and catalytic activity molecules; in biological process, mainly involved in cell proliferation, differentiation, apoptosis and metabolism. Meanwhile, the target genes of miR-376b-5p ( IGF-1, DAZL, MTOR, MET and NEDD4 ) and miR-376b-5p ( AHR ) take part in reproduction regulation.【Conclusion】Sheep ovarian miRNAs expression profiling was successfully constructed, and differentially expressed miRNAs between single-lamb-producing and twin-producing sheep were obtained. These miRNAs may be involved in sheep follicular development and litter size regulation.

【Objective】 This experiment was conducted to evaluate the effects of dietary protein sources on egg performance and egg quality of Jinghong laying hens at peak production. 【Method】 Two hundred and eighty-eight Jinghong laying hens aged 32 weeks with similar egg production were randomly divided into 4 groups with 6 replicates per group and 12 hens per replicate. Hens in group Ⅰ (control group) were fed soybean meal (SBM) protein, hens in group Ⅱ were fed degossypolized cottonseed protein (DCP), hens in group Ⅲ were fed double-zero rapeseed meal (DRM) protein, and hens in group Ⅳ were fed 4% yellow mealworms (YM) and soybean meal (SBM) protein. All diets were isocaloric (11.11 Mcal·kg^-1), isonitrogenous (CP, 16.5%) and had the similar standard ileum digestible AA model. The experiment lasted for 8 weeks. 【Result】 (1) In the whole feeding period，no significant differences were found on average egg production, egg weight and cracked eggs ratio among all groups (P＞0.05). In DCP group, egg mass during 1-8w was lower than that SBM group or 4% YM group (P＜0.05), while the egg mass of DRM group was lower than that SBM group（P=0.084）. The average feed intake of DCP group and 4% YM group during 1-8w were lower than that SBM group (P=0.091), while the average feed intake of DRM group was lower than that SBM group（P=0.091）. Egg/feed also showed that the same tendency in all groups, that is the egg/feed of DCP group and the DRM group were reduced (P=0.052), while the egg/feed of 4% YM group had no significant difference (P＞0.05). (2) No significant differences in the plasma aspartate transaminase (AST) among all groups were observed (P＞0.05). Compared to SBM group, plasma alanine transaminase (ALT) in laying hens of 4% YM group was significantly reduced at 8w (P＜0.05). The plasma total protein, globulin in laying hens of DCP group at 8w were significant lower than that SBM group (P＜0.05). The plasma albumen in laying hens of 4% YM group was higher than those DCP group and DRM group at 4w and 8w (P＜0.05). (3) There was a trend toward eggs of DCP group and DRM group at 8w decreasing shell strength (P=0.084) and shell thickness (P=0.062), while the egg of 4% YM group increasing shell strength (P=0.084) and shell thickness (P=0.062). Compared to SBM group, albumen height of eggs in DCP group and DRM group were reduced 4.02% and 3.41% (P=0.091) at 4w, while the 4% YM group was increased 3.87% (P=0.079). No significant difference were found albumen pH among all groups (P＞0.05). (4) Albumen percent of eggs in DCP group and DRM group were reduced 1.67% and 1.18% (P=0.099) than that SBM group at 4w, while 4% yellow mealworms group was increased 1.70% (P=0.099). In addition, yolk dry matter of egg in 4% YM group was higher than that DRM group at 4w (P＜0.05). 【Conclusion】 4% yellow mealworms replace soybean meal and soybean meal was the most optimal protein ingredients for Jinghong laying hens at peak production, followed by double-zero rapeseed meal and degossypolized cottonseed protein.

【Objective】 By using antimicrobial susceptibility testing and detection of phenotype and genotype in extended spectrum beta-lactamases (ESBLs)， this study aims to find out the distribution and drug-resistance of ESBLs-producing E.coli strains on broiler farms in Qingdao, and then analyze genotype and gene subtype of ESBLs, so as to provide a basis for guiding rational clinical use of antimicrobial agents and effective control of infection and spread of ESBLs-producing strains.【Method】The antimicrobial susceptibility of 249 E. coli strains was determined by micro broth dilution, the phenotype of ESBLs was identified and determined by CLSI standard method, DNA was amplified and the genotype of drug resistance plasmid of ESBLs was analyzed by PCR, sequencing and biological software, and the significant difference between ESBLs-producing and non-producing strains was analyzed  with SPSS19.0 software.【Result】There were 83.13% of E.coli strains produced ESBLs. The resistance rate of ESBLs-producing strains to 7 kinds of drugs was higher than that of non-ESBLs-producing ones, and there was a significant difference between them (P＜0.05) to GM, SPT, AM, A/C and CEF. But the resistance rate of ESBLs-producing strains to tetracycline and florfenicol was significantly lower than those in non-ESBLs-producing ones. The multi-drug resistance rate of ESBLs producing and non-producing strains was 99.03% and 92.86%, respectively (P=0.035). The detection rate of CTX-M, TEM and OXA genotype was 100%, 99.52% and 47.83%, respectively, and SHV genotype wasn’t detected. The enzyme producing strains belong to 10 gene subtypes, the dominant ones of which were TEM-1, CTX-M-65, CTX-M-55, and OXA-1. CTX-M-123 and CTX-M-64, the recombinant chimeras, were found from healthy poultry for the first time. 【Conclusion】The chicken origin ESBLs-producing E.coli strains are widely spread and transmitted in Qingdao. The antimicrobial resistance of ESBLs-producing strains are more serious than that of non-ESBLs-producing strains. Compared to other areas in China, CTX-M and TEM are also popular genotypes in Qingdao, but with different gene subtypes, TEM-1, CTX-M-65, CTX-M-55 and OXA-1 are dominant gene subtypes of each genotype, respectively.

【Objective】The objective of this study is to analyze the effect of Brassinosteroid (BR) on Ca²⁺distribution of plant cell, analyze the influence of BR on the expression level of the genes coding Ca²⁺ channel and Ca²⁺-ATPase, and clarify the effect of BR on calcium homeostasis in Arabidopsis thaliana. 【Method】The effect of BR on Ca²⁺ location in plant cell was studied using the potassium pyruantimonatc technique. The expression level of genes which coded Ca²⁺-ATPase (located in the membrane, vacuole, endoplasmic reticulum) and Ca²⁺ channel (located in membrane, vacuole, lysosome) was studied by Real-time PCR technique.【Result】Under normal condition, Ca²⁺ mainly distributed in cell walls, intercellular space, vacuole, and there were only a few Ca²⁺ distributed in the chloroplast and cytoplasm in plant cells. After 1 µmol·L^-1 BR treatment for 3 hours, Ca²⁺ gathered nearby vacuole membrane and cell membrane, meanwhile, the distribution of Ca²⁺ in the cytoplasm and chloroplast was increased. After BR treated for 6 hours, the Ca²⁺ distribution in the cytoplasm and chloroplast was continue increased, the Ca²⁺ distribution in the cell walls was reduced; after BR treated for 9 hours, the Ca²⁺ distribution in the cytoplasm and chloroplast was decreased, but increased in the intercellular space and vacuole, while the Ca²⁺ distribution in the cell walls was obviously reduced. It suggested that BR had function to release Ca²⁺ from cell wall. CNGC2 and CNGC12 are genes which encoded Ca²⁺ channel in the cell membrane. After 1 µmol·L^-1 BR treated for 3 hours, the expression level of CNGC2 and CNGC12 were all obviously decreased, this indicated that BR could block the extracellular Ca²⁺ transfer into cytoplasm. After 1 µmol·L^-1 BR treated for 6 hours, the expression levelsof CNGC2 and CNGC12 recovered；after 1 µmol·L^-1 BR treated for 9 hours, the expression levels of CNGC2 and CNGC12 were obviously increased. TPC1 and TPC2 are genes encoding Ca²⁺ channel in the vacuole and lysosome, respectively. The expression levels of TPC1 and TPC2 were significantly decreased after 1 µmol·L^-1 BR treated for 3 hours, but the TPC1 expression level was obviously higher than that without BR treated; the expression level of TPC2 was also significantly increased after 1 µmol·L^-1 BR treated for 9 hours. The results showed that BR could block the fast rising of the cytoplasm Ca²⁺ Concentrations, the expression level of the genes coding vacuole Ca²⁺ channel recovered earlier than that of the Ca²⁺ channel genes in the cell membrane and lysosome. ACA8 and ACA10 are genes encoding Ca²⁺-ATPase located in the cell membrane. After 1 µmol·L^-1 BR treated for 3 and 6 hours, the expression level of ACA8 and ACA10 had no significant change. After BR treated for 9 hours, the expression level of ACA8 and ACA10 was significantly increased. ACA4 and ACA11 were genes encoding Ca²⁺-ATPase located in the vacuole membrane, the change of ACA4 and ACA11 expression level was similar to the change of ACA8 and ACA10. ACA2 was one gene encoding Ca²⁺-ATPase located in the endoplasmic reticulum, the expression level of ACA2 also appeared the highest peak after 1 µmol·L^-1 BR treated for 9 hours. The results indicated that the expression level of Ca²⁺-ATPase genes increased after BR treated for 9 hours, and high concentrations cytoplasm Ca²⁺ was infused into extracellular and intracellular calcium store including intercellular space, vacuole, endoplasmic reticulum, etc. So BR could regulate the cytoplasm calcium homeostasis.【Conclusion】The second messenger Ca²⁺ was regulated by BR, and the signaling of BR was conducted through regulating the calcium homeostasis regulation system.

【Objective】Feeding deterrents or anti-feedants are a broad range of chemicals regulating the feeding behaviour via altering the palatability of crops to herbivores. The idea of pest ecological control using feeding deterrents is an alternative strategy to overcome the negative effect of chemical control. However, almost all insect pests can develop experience-dependent adaptivity to various feeding deterrents, especially gustatory habituation, which limited the practical use of these substances. The objective of this study is to elucidate the sensory organs involved in the feeding deterrence of two amides against a generalist herbivore, Helicoverpa armigera (Hübner), and the effect of prior feeding experience of piperine and sanshool on subsequent feeding behaviour of the larvae, providing a scientific basis for large-scale application.【Method】The olfactory approaching response under dual-choice condition with piperine- and sanshool-treated with tobacco leaf discs was tested, respectively, paired with the control leaf disc, and the feeding duration of the larvae after first encounter with the piperine- or sanshool-treated tobacco leaf discs under non-choice condition was measured. Then the effect of larval feeding experience to piperine and sanshool on their subsequent choice feeding behaviour was tested based on the concentrations at 50% feeding deterrence against the fourth instar larvae. 【Result】Piperine and sanshool did not exhibit any repellent effect, and the feeding durations of the larvae after first encounter with piperine- and sanshool-treated tobacco leaf discs were approximately 30 s, significantly shorter than that of the control (longer than 100 s). This feeding duration time was in accordance with the typical post-ingestive response time of a model insect species, Manduca sexta. The concentrations at 50% feeding deterrence to the fourth instar larvae of piperine and sanshool were 0.2259 and 0.4003 mg per tobacco leaf disc (1.5 cm ID), respectively. The bioassay of feeding experience showed that the larvae with piperine- and sanshool-feeding experiences were significantly deterred by the corresponding experienced substances, suggesting that no significant habituation occurred when the two amides were mixed with the artificial diet at the concentrations of 50% feeding deterrence to the fourth instar larva and when larval feeding experience was induced from the third instar to the fifth instar. The larvae experienced with sanshool were also deterred significantly by piperine. However, the larvae experienced piperine developed gustatory habituation to sanshool, demonstrating an asymmetric cross-habituation between these two amides. The binary mixture of piperine and sanshool could not mitigate the gustatory habituation of the larvae, whether considering the difference between the feeding consumption of the treated and control tobacco leaf discs within each experience group, or considering the feeding deterrence rates between of different experience group when tested under the same conditions. The possible reasons were discussed with respect to the induced stage and the mechanism of the tested substances deterring feeding. The binary mixture of piperine and sanshool could not mitigate the decrease in feeding deterrent response following prolonged exposure.【Conclusion】Feeding of H. armigera larvae was deterred by piperine and sanshool via gustatory post-ingestive effect. No significant gustatory habituation occurred when the experienced stimuli paired with the control, and it seems that the binary mixture of piperine and sanshool could not mitigate the habituation. The two amides have a broad prospect in field application.

【Objective】 Three long empty glumes mutants were screened from a maintainer line Yixiang 1B(Oryza sativa L. ssp. indica) by ethyl methanesulfonate (EMS) mutagensis. This study is to explore the genetic basis and molecular mechanism for controlling the development of long empty glumes by gene mapping and cloning. And the temporal and spatial expression of related genes were also concerned. 【Method】 Three long empty glumes mutants, Oslg-1, Oslg-2 and Oslg-3, were analyzed by phenotypic observation, allelism identification, gene mapping, bioinformatics analysis, and expression analysis by quantitative PCR. 【Result】The glumes showed no obvious difference at the stage of early floret differentiation between wild type and mutant. However, the epidermal cells of glume presented significant difference during floret maturation. At the heading date, epidermal cells of abaxial glume presented bulge and rough nodules, and formed axially alignment. The epidermal hairs became more, and bulges arranged at a regular intervals, which were similar to the structure of epidermal cells of lemma. Genetic analysis suggested that the mutant Oslg-1 was controlled by one recessive nuclear gene. The Oslg-1gene was mapped between SSR markers RM5344 and RM20934, on the short arm of rice chromosome 7, with genetic distances of 1.11 cM and 0.82 cM, respectively, and with a physical interval of 246.3 kb. By analyzing and sequencing for the candidate genes in this genomic region, it was found that there was a single nucleotide change at 182 base (T→A) in exon of LOC_Os07g04670 gene, which caused a missense mutation at 61 amino acid (Leu to His) in the encoded product. Allelism analysis by inter-crosses showed that Oslg-1, Oslg-2 and Oslg-3 were allelic variation. Detection of the gene OsLG in Oslg-2 and Oslg-3 mutants was made and found that there was a single nucleotide change (316, T→A and 119, T→C) in exon of OsLG gene, which caused a missense mutation (106, Trp to Arg and 40, Leu to Pro) in the encoded product, respectively. These results suggested that the candidate gene may regulate the elongation of rice glume by the homologous gene sequence alignment and phylogenetic analysis. Besides, the relative expression content involved in OsLG gene and another PAP2 genecontrolling glume trait was also analyzed by real-time PCR (qRT-PCR). The results showed that OsLG gene was expressed in roots, leaf sheath and panicle, and presented rather higher expression in panicle than that in root and leaf sheath. By contrast, PAP2 gene was only detected in panicle. Therefore, both of OsLG and PAP2 were tissue-specificaly expressed, and PAP2 was much more higher expressed in panicle than OsLG. It was deduced that these two genes functioned in a synergistic pattern to control the development of long empty glumes as the expression level of OsLG and PAP2 were down in OsLG mutants. 【Conclusion】The Oslgmutants are allelic with G1 reported previously, and the mutation of a single amino acid in functional domain leads to the development of long sterile glume. OsLG and PAP2 are synergistically expressed in panicle.