Table of Content

01 November 2015, Volume 48 Issue 21

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Characterization and Candidate Gene Analysis of Yellow-Green Leaf Mutant ygl13 in Rice (Oryza sativa)

WANG Ya-qin, SHI Jun-qiong, ZHANG Ting, LI Yan, ZHANG Tian-quan, ZHANG Xiao-long, SANG Xian-chun, LING Ying-hua, HE Guang-hua

Scientia Agricultura Sinica. 2015, 48(21):  4197-4208.  doi:10.3864/j.issn.0578-1752.2015.21.001

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【Objective】The current study was conducted aiming at phenotypic characterization and candidate gene analysis of the yellow-green mutant ygl13, so as to add to our knowledge of the formation and regulation of the molecular mechanisms responsible for leaf-color mutations in rice.【Method】A new rice mutant exhibiting stable inheritance was identified as derived from ethyl methane sulfonate (EMS)-treated restorer line Jinhui10 (Oryza sativa), tentatively named as yellow-green leaf 13 (ygl13). Morphological characteristics, the photosynthetic pigment contents and the agronomic traits were measured systematically. Transmission electron microscopy was conducted to analyze the ultrastructure of the mesophyll cells and chloroplasts in the ygl13 mutant and wide-type plants. The ygl13 was crossed with indica sterile line Xinong1A whose plant and leaves were normally green, and the morphological phenotype and segregation ratio of F₁ and F₂ were used for genetic analysis, F₂ for gene mapping, and putative genes in the fine mapped region were analyzed, and the candidate genes in the mutant and the wild type were sequenced, respectively.【Result】The ygl13 leaves displayed yellow-green compared with the wild type. And the photosynthetic pigment contents of chlorophyll a, chlorophyll b, and carotenoid decreased significantly at the seedling and the booting stages. The results from transmission electron microscope demonstrated that the structure of the chloroplast in the mutant ygl13 developed abnormally with poor thylakoids, less grana stacks and scattered distribution when compared with the wide type. According to the performance of agronomic traits, compared with the wild type Jinhui 10, the grain number per panicle increased by 26.06%, and the pant height and seed setting rate decreased by 12.33% and 18.82%. As for the panicle length, effective panicles per plant, filled grain number per panicle and 1000-grain weight, there was no significant difference between the wild type and ygl13. Genetic analysis demonstrated that the mutant trait was controlled by a single recessive gene as the number of green seedlings verses that of yellow-green seedlings approached 3:1（χ²=2.35＜χ²_0.05=3.84). Genetic mapping of the mutant gene was conducted using 602 recessive individuals from the F₂ segregation population. Finally, YGL13 was mapped on the short arm of chromosome 8 between InDel marker ID43 and ID69, and with an interval of 318 kb. There were 52 genes in this region and the sequencing analysis of these candidate genes between the mutant and its wild type revealed a single base change (G1005A) of the OsSIG1 gene (LOC_Os08g06630) in the encoded product resulted in a premature stop codon and protein truncation with 334 residues not the primary protein with 520 residues in the mutant ygl13. The qRT-PCR results showed that, the expression level of genes associated with pigment metabolism and photosynthesis is in disorder. 【Conclusion】 The mutant ygl13 was controlled by a single recessive gene. The gene YGL13 was allelic to OsSIG1 which was documented as a plastid sigma factor previously in rice. 

The Coding Regions Allelic Variations of TaGW2-6A and the Relationship Between the Allele and Drought Tolerance of Wheat (Triticum aestivum L.)

LI Yang, LI Li-qun, GAO Xin, YANG Lu, KOU Cheng, Lü Qian, LIU Tian-hong, DU Deng-feng, LI Xue-jun

Scientia Agricultura Sinica. 2015, 48(21):  4209-4218.  doi:10.3864/j.issn.0578-1752.2015.21.002

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【Objective】 Drought is one of the most significant factors that restrict wheat (Triticum aestivum L.) growth and reduce productivity. The kernel size and its stability affect the yield of wheat in different rainfall locations. The research of grain weight and the relationship between the drought resistance has a vital significance to high and stable yield. The aim of this study is to gain further insight into the drought resistance of different allelic variations of kernel weight related gene, determine the allelic gene variant genotype with strong drought resistance and high yield ability, making for marker-assisted selection of drought resistance and high yield varieties and provide a theoretical basis for genetic improvement. 【Method】 An allelic variation with single T-base insertion at 977 bp of TaGW2-6A was detected in Lankaodali with large kernel, designated as TaGW2-6A-T (TT genotype), but was not found in Chinese Spring (tt genotype). According to the sequence of “T” insert, primers were designed to detect the coding regions allelic variations of TaGW2-6A by High Resolution Melting Curve (HRM) analysis technique, and the sequence of PCR products were analyzed. To gain further insight into the role of allelic variations of TaGW2 in drought tolerance, various physiological parameters related to stress response and kernel weight related traits including the chlorophyll fluorescence kinetics parameters, stem soluble sugar content, drought resistance index and kernel weight related traits of 325 recombinant inbred lines (RILs) derived from a cross between Chinese Spring and Lankaodali were analyzed under different water conditions at flowering, filling and milk stages.【Result】 Results showed that it was accurate to detect the wheat coding regions allelic variations of TaGW2-6A by HRM analysis technique, and classified as Lankaodali genotype (TT), Chinese Spring genotype (tt) and heterozygous genotype (Tt). The PCR product sequencing results showed that the solubility curve difference was caused by the “T” base insert. Compared to tt genotype, the kernel length, kernel width and kernel weight of TT/Tt genotype increased significantly. The chlorophyll fluorescence kinetics parameters, drought resistance index and stem soluble sugar content of the two genotypes were significantly or extremely significantly different (P＜0.05 or P＜0.01). The chlorophyll fluorescence kinetics parameters of RILs and their parents reduced, except the initial fluorescence Fo, while the stem soluble sugar content increased under drought stress. The variability of chlorophyll fluorescence parameters of RILs with TT/Tt genotype was smaller than that of RILs with tt genotype. While stem soluble sugar content of RIL with TT genotype was higher than that of RILs with tt genotype in Yangling and Qianxian. The accumulation efficiency and remobilization efficiency of stem soluble sugar content increased with TT/Tt genotype, while decreased with tt genotype. In addition, TT genotype plays an important role to the chlorophyll fluorescence parameters, the stem soluble sugar content and kernel weight. The study revealed that, compared with the RILs with tt genotype, the photosynthetic institutions of RILs with TT genotype damaged smaller, and accumulated more osmotic regulation substances to ensure the grain filling when suffering drought stress. 【Conclusion】 TaGW2-6A variation (TT genotype) was better able to prevent reducing production in different locations with strong drought resistance and stable yield ability than wild-type (tt) plants, and the ability of drought resistance is close to Jinmai 47.

Cloning and Expression Analysis of the Potato Transcription Factor StWRKY8 Like Gene Induced by Ralstonia solanacearum

XUE Zhen, LI Hui, KONG Chao-yue, DUAN Ting-ting, GAO Gang

Scientia Agricultura Sinica. 2015, 48(21):  4219-4226.  doi:10.3864/j.issn.0578-1752.2015.21.003

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【Objective】Clone the partial cDNA of the StWRKY8-like gene from a cultivated Solanum tuberosum Zhongshu 3 after inoculation with Ralstonia solanacearum. Analyze the StWRKY8-like gene coding sequence, and study the differential expression pattern of StWRKY8-like between susceptible and resistant S. tuberosum toward R. solanacearum and the tissue-specific expression of StWRKY8-like.【Method】R. solanacearum-resistant ED13 and R. solanacearum-susceptible Zhongshu 3 S. tuberosum were root-inoculated with R. solanacearum strain PO41. RNA were extracted from the leaves and reserve-transcribed into cDNA which was subjected for the construction of subtractive cDNA bank with the PCR Select cDNA Subtraction Kit. 384 positive clones were obtained as SMART cDNA bank and used as a template for 5′-RACE with SMART-RACE cDNA Amplification Kit to PCR amplify the StWRKY8-like gene. The StWRKY8-like gene sequence and DNA sequence similarity were analyzed by BioEdit and BLAST tools. Phylogenetic trees were established by MEGA 5.0. The biochemical feature, tertiary structure, phosphorylation sites and sub-cellular location of the StWRKY8-like protein were predicted by ProtParam/ProtScale, SWISS-MODEL, NetPhos2.0 Server, WOLF PSORT/TargetP1.1 Server, respectively. The RNA were extracted from the leaves of ED13 and Zhongshu 3 S. tuberosum at 6 h, 12 h, 1 d, 2 d, 3 d, 4 d, and 6 d post inoculation of the R. solanacearum strain PO41 for RT-PCR and real-time PCR analysis of the StWRKY8-like gene expression. Digoxin-labeled StWRKY8-like specific probes were in situ hybridized with S. tuberosum stem and leaf sections to determine its expression in different tissues. 【Result】 The partial cDNA of the StWRKY8-like gene (563 bp) was obtained, which contained a 258 bp open reading frame coding a peptide with 85 amino acids. The StWRKY8-like protein has a classical conserved WRKY domain with a zinc finger motif of C₂H₂ and belongs to the subgroup II of WRKY family. The amino acid sequence of StWRKY8-like protein was highly close to the other WRKY members of Solanaceous plants and in 98% similarity with StWRKY8 in R. solanacearum. The StWRKY8-like protein is predicted as a hydrosoluble protein with an isoelectric point of 9.1 and half-life period of 5.5 h. It is a non-spherical protein, contains 3 phosphorylation sites and locates in the cytoplasm. The StWRKY8-like gene was upregulated after the R.solanacearum infection, and differentially expressed in susceptible and resistant S. tubosum. The expression of StWRKY8-like in resistant S. tubosum was significantly higher than in susceptible S. tubosum at 6 h post infection. It was mainly expressed in the vascular system of the stem and leaves. 【Conclusion】 StWRKY8-like was a putative transcription factor gene, upregulated at R. solanacearum infection and differentially expressed in susceptible and resistant S. tubosum. It may be involved in the S. tubosum defense against R. solanacearum.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY

Effect of Auxin Treatment on Root Hair Formation and Aquaporins Genes Expression in Root Hair of Rice

MO Yi-wei, LI Xia-jie, WANG Hai, CHEN Ze-kai, YANG Guo, WANG Wei

Scientia Agricultura Sinica. 2015, 48(21):  4227-4239.  doi:10.3864/j.issn.0578-1752.2015.21.004

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【Objective】To explore the effect of auxin (IAA) and its polar auxin transport carriers on the formation of rice root hair and genetic expression of aquaporins.【Method】The varieties of wild-type Japonica rice Zhonghua11 and its over-expressed OsPIN1a transgenic rice primary root were used in this paper. A small agar block which contains different concentrations of IAA and combination of IAA and inhibitors of polar auxin transport efflux carrier TIBA (2, 3, 5-Triiodobenzoic acid), NPA (N-1-naphthylphthalamic acid) or polar auxin transport influx carrier CHPAA (3-Chloro-4-hydroxyphenylacetic acid) was affixed to the lateral side of rice root tip when the primary root reached 0.5—1.0 cm in length in a aeroponic and dark culture. After a 12h treatment, the root hair length, root growth rate, root hair density and relative water content in rice roots were detected. The photos were taken by confocal laser scanning microscope (Zeiss Axiophot microscope with a Qimaging Retiga 1300 12-bit monochrome CCD camera with Open Lab v3.0.9) to observe subcellular localization of OsPIN1a-GFP in the tip and hair of the rice root. In addition, semi-quantitative RT-PCR was applied to detecting the OsPIN1a and aquaporins genes expression before and after the formation of the rice root hair. 【Result】The results showed that the length and density of the root hair enhanced correspondingly with an increase in IAA concentration range from 0 to 5.0 mg·L^-1, and 2.5 mg·L^-1 IAA had an optimal effect on inducing the formation of root hair. However, as IAA exceeds 5.0 mg·L^-1, the increase in root hair was insignificant and thus the primary root growth was severely inhibited. The new root hair only appeared when agar was attached to the meristematic zone of root tip but not formed in the ripe areas of the primary root. Combined treatment with IAA and lower concentration of TIBA or NPA did not affect formation of root hairs. In contrast, higher concentration of TIBA or NPA significantly inhibited its formation and elongation. However, much lower concentrations of CHPAA can significantly inhibit the formation and elongation of the root hair. In addition, IAA treatment increased genes expression of aquaporins as well as the formation of the root hair, thus improving the relative content of water in root tips. IAA treatment markedly improved the expression of OsPIN1a and the content of OsPIN1a-GFP in root tips and root hair.【Conclusion】It was suggested that the formation of rice root hair was induced by IAA and associated with the involvement of IAA efflux and influx transporters and influx process had the most significant effect. OsPIN1a played a key role in the formation and elongation of the root hair. IAA can promote genes expression of aquaporins and increase relative water content, thus alleviating water stress in rice roots in the aeroponic culture.

Comparison on Morphological, Yield, and Quality Traits of Soybean Cultivars Developed in Different Years from Liaoning and Ohio

GUO Xiao-hong, WANG Xing-cai, MENG Tian, ZHANG Hui-jun, AO Xue, WANG Hai-ying, XIE Fu-ti

Scientia Agricultura Sinica. 2015, 48(21):  4240-4253.  doi:10.3864/j.issn.0578-1752.2015.21.005

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【Objective】The research compared the morphological, yield, and quality traits of soybeans [Glycine max (L.) Merr.] cultivars developed in different years from different breeding programs of Ohio, U.S.A. and Liaoning, China, which were at the same latitude. The purpose of this study was to reveal the evolutionary trend of the main agronomic traits, and the response to the fertilizer rate with the seed yield improvement.【Method】The field experiment was conducted in 2011, 2013, and 2014, and used a completely randomized design with three replications. The diammonium phosphate was applied as starter fertilizer, and had three levels (0, 150, and 300 kg·hm^-2). The main agronomic traits, seed yield, and quality were measured at the mature stage.【Result】With the improvement of the soybean yield of Liaoning and Ohio cultivars, plant height, internode length, and lodging score decreased, harvest index increased; the branch and height of the lowest pod of Ohio cultivars increased, and that of Liaoning cultivars was decreased. The improvement of seeds per plant was the main reason for the yield increasing of Ohio cultivars, and the main factor for Liaoning cultivars was of a bigger 100-seed weight. In the process of cultivar development, the protein content of Ohio cultivars decreased, while the fat content increased; the protein content of Liaoning cultivars increased, and there was no significant changed in fat content. Compared with Liaoning cultivars during the same period, the plant height, internode length, and lodging score of Ohio cultivar decreased; and pods per plant, seeds per plant, harvest index, and yield increased; protein content decreased, and fat content increased. With the increment of the fertilizer rates, the plant height, internode length, and lodging score of the Ohio cultivar remained relatively stable, but that of Liaoning cultivars increased sharply; the yield of Ohio cultivars increased with the fertilizer rates, and the yield of Liaoning cultivars was highest at the medium fertilizer rate. The results of correlation and path-coefficient indicated that the yield was significant negatively correlated with plant height, internode length, and lodging score; while it was significant positively correlated with seeds per plant and harvest index, and plant height, seeds per plant, and harvest index had a greater direct effect to yield improvement.【Conclusion】During the improvement of the soybean yield of the Ohio and Liaoning cultivars, the plant height and internode length decreased, the lodging resistance and harvest index increased, but the improvement progress of the Ohio cultivars was greater than that of the Liaoning cultivars. The yield improvement of the Ohio cultivars was mainly attributed to the increment of seeds per plant, and that of Liaoning cultivars was a bigger 100-seed weight. The fertilizer tolerance of the Ohio cultivars was enhanced, and the plant height and lodging score kept stable by the fertilizer rates rising, but that of the Liaoning cultivars increased.

Effects of Symbiotic Period on Seedling Traits and Yield Components of Interplanting Rapeseed in Rice

ZHENG Wei, YE Chuan, XIAO Guo-bin, CHEN Ming, LI Ya-zhen, HUANG Tian-bao, XIAO Xiao-jun, LIU Xiao-san, ZHU Chang-lan

Scientia Agricultura Sinica. 2015, 48(21):  4254-4263.  doi:10.3864/j.issn.0578-1752.2015.21.006

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【Objective】The main purpose of this study was to recover the optimal strategy for growth area expansion and output increasing of interplanting rapeseed in rice, a triple cropping pattern, through evaluating the effects of a symbiotic period on both seedling growth and yield development, which would shed light on desirable symbiotic period determination in South China.【Method】Ganyouza 5 and Fengyou 730, two elite rapeseed cultivars, were employed for a field experiment with 6 symbiotic period treats during the 2012-2014 growing season, the 6 symbiotic period treats were as followed separately: 12 d, 10 d, 7 d, 5 d, 3 d, 0 d (CK).【Result】(1) An extended symbiotic period could thoroughly promote the seedling density. A dramatic reduction of seedling density occurred along with a longer symbiotic period in the late growth period. Concurrently, the seedling rate and mature plant rate were increased at the initial stage but deteriorated under a further extended symbiotic period. The optimal mature plant rate was a 5-day symbiotic period treatment. Compared with the control (0-day symbiosis), the seedling rate and mature plant rate were raised with 0.97% and 5.98% at a 5-day symbiotic period but lessened by 19.2% and 19.35% at a 12-day symbiotic period, respectively. (2) Under a longer symbiotic period, seedling growth, number of total leaves, number of green leaves, crown diameter, and the dry weight of individual plants were gradually enhanced, whereas the leaf area index and dry matter of population plants were primitively increased and then subsequently fell. (3) A symbiotic period was highly related to yield components of interplanting rapeseed in rice including primary branches, siliques per plant, siliques on main inflorescence, seeds per silique, 1000-grain weight, dry matter weight per plant, and yield per plant. Especially, a significant positive correlation was observed between the symbiotic period and the primary branches, siliques per plant, siliques on main inflorescence, 1000-grain weight, yield per plant, and dry matter weight per plant, but a substantial negative correlation occurred for seeds per silique. (4) The yield per plant of interplanting rapeseed in rice was enhanced, whereas the yield per area was heightened initially and diminished later under a longer symbiotic period, and the optimal rapeseed yield could be approached at 5-day symbiotic period. Comparing with the control (CK), the average yield increasing rate of the symbiotic period treatment was 20.48% and 22.35% respectively within two rounds of field experiments. (5) The duration of a symbiotic period could affect the developing progress of rapeseed. The life period of the control (0-day symbiosis) was shorter than the symbiotic treats, and the difference was apparently occurred from seedling to bolting stages, but disappeared after the squaring stage.【Conclusion】Extension of a symbiotic period favors seedling density and individual development of interplanting rapeseed in rice. However, a lower seedling rate and mature plant rate under a longer symbiotic period as well as low seedling density and weak individuals under a shorter symbiotic period could result in yield reduction. Our results indicated that 5-days was the most optimal local symbiotic period within a favorable regime of 3 to 7 days to higher yield under interplanting rapeseed in rice in red soil regions. A longer or shorter symbiotic period would be detrimental to high rapeseed yield formation in South China where a triple cropping system is very popular.

PLANT PROTECTION

Effect of Mixed Infection of Cercospora arachidicola and Phoma arachidicola on their Infection Probability and Latent Periods

ZHOU Ru-jun, CUI Jian-chao, FU Jun-fan, XU Zhe, XUE Cai-yun

Scientia Agricultura Sinica. 2015, 48(21):  4264-4271.  doi:10.3864/j.issn.0578-1752.2015.21.007

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【Objective】Early leaf spot disease and web blotch disease are the most serious disease in peanuts. The symptoms of these two diseases always occur on the same plant in the late growth stage of the peanut, and cause serious yield losses in peanut production. The objectives of this study are to evaluate the factors that influence the infection probability and latent period of Cercospora arachidicola and Phoma arachidicola after mixed infection on peanuts, investigate the interactions between disease and host plants, and provide a basis for formulating comprehensive control of more diseases and pests on the same host.【Method】C. arachidicola and P. arachidicola were inoculated on susceptible cultivar, Baisha1016. In the field, different plant growth stages, inoculum concentration and leaf wetness period were constructed. The influences of the mixed infection of C. arachidicola and P. arachidicola on the infection probability and latent period were analyzed.【Result】When C. arachidicola and P. arachidicola were inoculated alone, the infection probability of both pathogens were increased with the extension of the growth stage. Specifically, the infection probability for the different growth stage of peanut was: early flowering stage C. arachidicola and P. arachidicola were inoculated alone at different stages, there were significant differences between the latent period of the two pathogens. In the early flowering stage and late flowering stage, the latent period of C. arachidicola was 20 d, but in the flowering stage, the latent period of C. arachidicola was 16 d; in the early flowering stage, the latent period of P. arachidicola was 10 d, however, in the flowering stage and the late flowering stage, the latent period of P. arachidicola was 7 d. The latent period could be affected by leaf age and the degree of senescence, it was longer in the early growth stage than the late growth period, but had little significant correlation with temperature and humidity. Mixed inoculation of the two pathogens prolonged the latent period. In the early flowering stage, the latent period for C. arachidicola and P. arachidicola increased for 2 d and 5 d, compared to the pathogens inoculated alone. And in the flowering stage and the late flowering stage, the latent period for C. arachidicola and P. arachidicola increased for 4 d and 2 d, respectively, but the latent period of P. arachidicola had no obvious change, all were 7 d.【Conclusion】When the C. arachidicola and P. arachidicola were inoculated individually, the infection probability of the two pathogens increased with the extension of the growth stage and leaf wetness duration and the increase in inoculation concentration. However, when the two pathogens were inoculated simultaneously, the infection probability was decreased and the latent period was prolonged. In addition to the influence of the interactions between the pathogen and host, the latent period of C. arachidicola and P. arachidicola could be affected by leaf age and the degree of senescence, but had little significant correlation with temperature and humidity.

Bioinformatics and Tissue-Specific Expression Analysis of Carboxylesterase Genes from Oxya chinensis

LIU Jiao, ZHANG Jian-zhen, LI Da-qi, ZHANG Ting-ting, MA En-bo, ZHANG Jian-qin

Scientia Agricultura Sinica. 2015, 48(21):  4272-4284.  doi:10.3864/j.issn.0578-1752.2015.21.008

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【Objective】Oxya chinensis is an important agricultural pest. The objective of this study is to reveal the function of carboxylesterases (Ces) genes and select potential molecular target for pest management through conducting the bioinformatics and tissue-specific expression analysis of Ces genes from O. chinensis transcriptome. 【Method】 The cDNA sequences of Ces genes were searched using the keyword “carboxylesterase” against the O. chinensis transcriptome database. The Ces cDNA sequence assembling, the deduced amino acid sequence analysis and the open reading frame finding were performed by using bioinformatics methods. Phylogenetic tree of O. chinensis carboxylesterase with complete ORF was constructed using neighbor-joining method. The structure, calculated molecular mass, isoelectric points, signal peptide and potential N-glycosylation sites of O. chinensis Ces amino acid sequences were predicted using corresponding online softwares. Optimal reference genes in different tissues of 5th instar nymphs were screened and relative expressions in different tissues of 10 Ces genes were detected by reverse transcription- quantitative PCR (RT-qPCR).【Result】 A total of 180 putative Ces cDNA fragments were identified from O. chinensis transcriptome. Among them 28 full-length cDNAs containing complete ORFs were obtained. Phylogenetic tree of Ces protein sequences from O. chinensis and other insect species showed that 28 O. chinensis Ces protein sequences were distributed in four clades, including clade A with Orthopteran and part of Dipteran α-esterases (20 Ces protein sequences), clade D with integumental esterases (2), clade E with β-esterases (4), and clade F with nonlepidopteran JHEs (2). With the exception of three Ces proteins with alkaline isoelectric points (pI), all the remaining Ces proteins are acidic or slightly acidic with pI ranging from 4.38 to 6.84. All the Ces protein sequences have N-glycosylation site, N-terminal conserved cysteine residues, and oxyanion hole. Signal peptide prediction showed 19 of 28 have signal peptide. Twenty-one Ces protein sequences have conserved catalytic triad S-E-H, however, the remaining 7 Ces protein sequences showed different amino acid substitutions. GeNorm and Normfinder analysis showed that EF1α and RP49 were the optimal reference genes. RT-qPCR showed that 10 Ces genes were expressed in 7 different tissues, in which OcCesA1, OcCesA6, OcCesA12, OcCesA14, OcCesA18 and OcCesA19 mainly expressed in the midgut and gastric caecum; OcCesA6, OcCesA18, OcCesA19 still expressed in Malpighian tubules; OcCesA2 highly expressed in the hindgut. OcCesD1 had the highest expression in Malpighian tubules, followed by the fat body and integument. OcCesE1 had the highest expression in gastric caeca, OcCesF2 had the highest expression in midgut.【Conclusion】A total of 28 Ces cDNAs with complete ORFs were identified from the O. chinensis transcriptome database. These Ces cDNAs were clustered into four different clades in the phylogenetic analysis. Significant expansions of the Ces genes were found in clade A and highly expressed in detoxification, excretory organs, such as midgut, gastric caeca and Malpighian tubules. These results contributed to the further functional analysis of O. chinensis Ces and biological control of O. chinensis based on Ces gene molecular target.

SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT

The Ecological Development Potential and Time-Space Analysis of Chinese Main Agricultural Crop Biomass Energy

ZHU Kai-wei, LIU Zhen, Lü Zhi-chen, PU Gang-qing, GUO Wei

Scientia Agricultura Sinica. 2015, 48(21):  4285-4301.  doi:10.3864/j.issn.0578-1752.2015.21.009

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【Objective】The estimation of agricultural biomass resource is the key link and foundation to develop and utilize agricultural biomass scientifically. Straw returning could help maintaining and enhancing soil function, so the ecological potential assessment of agricultural biomass resource is the important prerequisite of sustainability using agricultural biomass. Thus, the main purpose of this article is, to consider the ecological requirements of agricultural soil, to assess the ecological available biomass energy potential from the main crops in different straw returning rate scenarios, and analyze the time-space characteristics and structural composition, to provide some suggestions for the development of agricultural biomass.【Method】Firstly, according to the national datum of crop planting area datum, the percentage ratio of each province and sown area, the main crops planting proportion and yield datum, using a linear regression method, expert prediction, to make predictions for the future 50 years; Secondly, based on the relative researches, the grass valley industrial using ratio, feed using ratio and burning ratio were designed; considering the soil ecological requirements and relative researches, using scenario analysis method, low crop remaining returning ratio scenario, medium crop remaining returning ratio scenario and high crop remaining returning ratio scenario were designed; Finally, from the perspective of spatial distribution and time distribution, to calculate and analyze the finally potential available biomass, and give some suggestions.【Result】(1) By 2050, in low crop remaining returning ratio scenario, medium crop remaining returning ratio scenario and high crop remaining returning ratio scenario, the final available biomass were respectively 162 million tons of standard coal equivalent, 78 million tons of standard coal equivalent and 11 million tons of standard coal equivalent. And as time goes on, the final available biomass of main crops will eventually have agglomeration on space, time and species composition. (2) In low and medium crop remaining returning ratio scenario, the space distribution of final biomass are mainly distributed in the northeast, East China, central China, besides Henan, Heilongjiang, Shandong and Xinjiang Province account most of the total; From the time aspect, it is mainly concentrated in August, September and October, and the rice, wheat and corn are the main sources. (3) In high crop remaining returning ratio scenario, from the space aspect, it mainly concentrated in East China, central China, South China and southwest China, besides Yunnan, Guangxi, Henan and Heilongjiang Province account most of the total; From the time aspect, it is mainly in January, February, November and December, from the component aspect, it is only made up of wheat, beans, potato and sugar cane.【Conclusion】Henan Province, Heilongjiang Province, Xinjiang Province and Shandong Province should be given priority when developing and using the biomass energy; Besides, when considering to build a regional crop straw distribution center, regional biomass energy industry base, large straw direct combustion power generation base, and some other large scale integrated agricultural biomass energy development projects, northeast China and central China could be given priority too. In addition, the biomass energy potential of southwest China, northwest China can be furthered through crop planting structure adjustment and improve the level of agricultural management.

Analysis on Process and Difference of Cropland Dynamics in Anji County of Zhejiang Province

CHEN Xue-yuan, TANG Hua-jun, WU Yong-chang, ZHOU Qing-bo, CUI Jian

Scientia Agricultura Sinica. 2015, 48(21):  4302-4313.  doi:10.3864/j.issn.0578-1752.2015.21.010

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【Objective】Land resource is a carrier of social economic activities, cropland is the link and bridge between human and environment, also is the most important part of the land system. It is helpful to improve the capacity to predict cropland change by understanding and mastering cropland dynamic change process and difference, and it is also beneficial to promote the scientific decision-making and intensive utilization of cropland, thus making agricultural land resource sustainable and social economy developing harmoniously.【Method】The research principally adopted the techniques of remote sensing and GIS, and used land use dynamic degree model, geographical econometric model including composite index of the degree of land use extension and transfer matrix of land use for the analysis of cropland dynamic change and difference in Anji of Zhejing Province in the two periods of 1998 to 2003 and 2003 to 2009. 【Result】The dynamic change process of cropland in research area was complex. The main types of land use transfer were irrigated land and arid land. From the point of types transfer, the main directions of cropland were forest land, construction land and garden land; the process of cropland transfer from 2003 to 2009 was smoother and steadier than the process from 1998 to 2003, and no dramatic changes were occurred. The types and area of the land resource which converted to cropland was very unitary and the area was also limited. All the types of land in the quantity and spatial distribution from 1998 to 2003 were stabler; the magnitude of cropland changes from 2003 to 2009 was lower in activity, so after 2003, cropland in the extent of utilization and range was stabler. 【Conclusion】The cropland dynamic change process and its characteristics are influenced directly by industry structure and land policy adjustment in the research area. Research methods used in this paper are operable and practical which have reflected the cropland dynamic change process and characteristics of space-time structure objectively, and have provided an idea for space-time evolution research of cropland.

HORTICULTURE

A Genome-Wide Survey of HD-ZipⅠ Genes and Their Responses to Hormone in Apple

WEN Xiao-hong, JIANG Yong-hua, KANG Jian, WANG Hao-jie,REN Xiao-lin

Scientia Agricultura Sinica. 2015, 48(21):  4314-4322.  doi:10.3864/j.issn.0578-1752.2015.21.011

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【Objective】By integrating bioinformatics and analyses of physiological features, the functions of homeodomain leucine zipper I (HD-Zip I) in apple genome were studied. This study may aid in the selection of appropriate candidate genes for apple ripening research.【Method】ExPASy was employed to analyze the family members’ basic physical and chemical properties, MEME and PLACE were used to predict cis-elements in the promoters and their functions, and GSDS was used to draw the stucture map of intron/extron. The ripening characters such as internal ethylene concentration, soluble solids, hardness and titratable acid after various hormone-treatments were tested. Semi-quantitative PCR was used to detect gene responses to different plant hormones.【Result】A total of 22 HD-Zip I genes were identified in Royal Gala, and they are further divided into ?ve clades with phylogenetic analysis. The intron/exon structures showed a partly conservation in structure from the same cluster. Cis-elements related to hormone responses (such as gibberellin, cytokinins, jasmonic acid, ethylene and abscisic acid) are widely distributed in the -600 bp upsteam region from ATG, and the most conserved and abundant one is (GA/TC)8 repeated sequence. The treatments of ethylene and abscisic acid similarly increased the inner ethylene content and promoted the maturation process. Differing from other members, the transcriptional levels of MdHZ1 and MdHZ17 were both up-regulated on 48 h after treatments, while their levels were down-regulated by PUT and GA4, which could delay the ripening process of apples in the experiments. These data showed a close connection between genes MdHZ1, MdHZ17 and fruit ripening. At the same time, it was also detected that the family members answered the hormone treatments differently and MdHZ16 was the undetectable one. Therefore, the up-regulate network of this family is complex and even the similar structure genes within a same family do not present alike expression patterns.【Conclusion】There are 22 HD-Zip I genes identified from apple and they possess similar gene structures. Promoter regions contain lots of cis-elements involved in hormone interaction response, and this may be a reason for the differently answer to exogenous hormone treatments. MdHZ1 and MdHZ17 could be the main candidate genes participating in ripening progress.

Study on Floral Scent of the Genus Hosta

LIU Qian, SUN Guo-feng, ZHANG Jin-zheng, LI Xiao-dong

Scientia Agricultura Sinica. 2015, 48(21):  4323-4334.  doi:10.3864/j.issn.0578-1752.2015.21.012

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【Objective】This paper evaluated floral scents and scent components in the genus Hosta in the germplasm in order to clarify the material basis of floral scent formation, which would provide necessary theoretical basis for further breeding efforts.【Method】This study was undertaken to 1) detect the floral scent components of 11 species and 129 cultivars in Hosta by headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS); 2) to investigate the variation in the fragrance patterns by sensory evaluation of their floral scents; and 3) to explore the origin of character aromas coupled with their genetic relationships.【Result】The floral scents of 130 hostas could be divided into seven aroma levels: not detectable (0), very low (0-1), low (1), low to medium (1-2), medium (2), medium to intense (2-3) and intense (3). Most (89%) hostas (first four levels) did not present distinct fragrances, and few hostas (latter three levels) produced strong sweet fragrances displaying a limited diversity of fragrance patterns. Large variation existed in the composition and contents of scent components emitted from Hosta flowers Fifty-nine volatiles were identified as scent components, including 38 terpenoids, 8 phenylpropanoids, 12 aliphatic compounds, and 1 nitrogenous compound. Terpenoids were the dominant components of these scents, composing between nearly 3/4 to all detected compounds. Large variation existed in the composition and contents of scent components emitted from Hosta flowers. Hosta plantaginea originating from China and its cultivated variety H. plantaginea ‘Aphrodite’ had the highest total emission contents of scent components as well as sum of terpenoids, phenylpropanoids, aliphatic compounds and nitrogenous compounds, corresponding to the most heavy and special scents emitted by a taxa. According to principal component analysis, the scent components of hosta flowers with strong sweet fragrances (medium, medium to intense and intense) notably differed from those without distinct fragrances (not detectable, very low, low and low to medium) due to their high emission contents of linalool. We define linalool to be the character impact compound, and its dominant synthesis leads to the strong sweet scent of hosta flowers. Myrcene, limonene, (E)-β-ocimene, in addition to linalool were mainly responsible for the formation of floral scents. This indicates the limited diversity of fragrance patterns in Hosta is due to a lack of high levels of aromatic compounds with low threshold values. Most hostas with strong sweet fragrances stem from H. plantaginea by hybridization or tissue culture, and others were possibly derived from the Japanese garden hybrid.【Conclusion】Hosta floral scents were divided into seven aroma levels. Terpenoids were the dominant components of hosta floral scents, among which myrcene, limonene, (E)-β-ocimene and linalool were mainly responsible for the floral scent formation. Linalool was identified as the character impact compound. The characteristic sweet scent derived mostly from lineages of H. plantaginea originating from China. These taxa are suggested to be the most valuable plant parent in Hosta with respect to scents. The exploration and application of other fragrant resources in Hosta require further study.

STORAGE·FRESH-KEEPING·PROCESSING

Effects of Ca²⁺ and Na⁺ Ions on the Structure and Rheological Property of Sugar Beet Pectin Under High Hydrostatic Pressure

PENG Xiao-yan, MU Tai-hua, ZHANG Miao, SUN Hong-nan, YU Ming, HE Wei-zhong

Scientia Agricultura Sinica. 2015, 48(21):  4335-4346.  doi:10.3864/j.issn.0578-1752.2015.21.013

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【Objective】 This study was in order to discuss the influence of metal ions (Ca²⁺ and Na⁺) sodium on the structure and rheological properties of sugar beet pectin under high hydrostatic pressure (HHP), and to provide theoretical basis for the application of sugar beet pectin in food. 【Method】 1% (w/v) sugar beet pectin solution which was prepared by 0.05 mol·L^-1 Tris - HCl solution with adding different concentrations of Ca²⁺ (2, 12 and 20 mmol·L^-1) and Na⁺ (0.05, 0.1 and 0.6 mol·L^-1), was treated under HHP, and the molecular weight, microstructure, viscosity and dynamic viscoelasticity of sugar beet pectin were investigated. 【Result】 Compared with the control one, new peaks were shown at 1 550 cm^-1 and the yield stress σ₀ increased significantly for sugar beet pectin treated at 450 MPa for 10, 20, 30 and 50 min, but there were little changes between the different treatment time. Sugar beet pectin with adding different concentrations of calcium or sodium ions was treated under 450 MPa for 30 min; the changes of its structure and rheological property were different. Relative to no add calcium or sodiumions, the σ₀, G’ and G” of sugar beet pectin increased significantly when adding 2 mmol·L^-1 calcium ions, when the concentration of calcium ions increased to 12 mol·L^-1 and 20 mol·L^-1, there was little change compared with add 2 mmol·L^-1 calcium; adding 2 calcium ions in the sugar beet pectin made the pectin crosslinking, and the molecular weight of sugar beet pectin was increased from 2.25 × 10⁵ Da to 6.07 × 10⁵ Da; with the concentrations of calciumions was increased to 20 mmol·L^-1, the molecular weight of pectin into 5.99 × 10⁵ Da, and there was no significant differences with the sugar beet pectin add 2 mmol·L^-1 calcium ions, and also its rheological properties. Relative to no add calcium or sodium ions, add 0.05 mol·L^-1 sodium ions also made the yield stress σ₀ of sugar beet pectin increase significantly, and with the increase of sodium ion concentration, yield stress σ₀ of pectin was increased significantly. when the concentration of sodium ion was increased to 0.6 mol·L^-1, the G’ and G” of sugar beet pectin was increased obviously, After adding 0.1 mol·L^-1 sodium ions, sugar beet pectin chains formed reticular by crosslinking with molecular weight significantly increased to 11.95 × 10⁵ Da. However, when added 0.6 mol·L^-1 sodium ion, the molecular weight of pectin was decreased to 5.53×10⁵ Da with virgate structure. 【Conclusion】 The calcium and sodium ions might change the structure of sugar beet pectin under HHP treatment, thus affect its structure and rheological properties.

ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT

Effect of Ammonia in Broiler Houses on the Antioxidant Activity and Meat Quality of Broiler

XING Huan, LUAN Su-jun, SUN Yong-bo, SA Ren-na, ZHANG Hong-fu

Scientia Agricultura Sinica. 2015, 48(21):  4347-4357.  doi:10.3864/j.issn.0578-1752.2015.21.014

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【Objective】The aim of this experiment is to study the effects of different concentration of ammonia on the antioxidant capacity and the meat quality of broiler chickens. 【Method】 The study has adopted a single factor completely randomized design. A total of 400 21-day-old Arbor Acres (AA) male broilers were randomly divided into 4 respiratory chambers, and each chamber was a treatment with 4 replicates, and 25 broilers in each replicate. The following concentrations of ammonia were set in different groups: ＜3 mg·kg^-1 (control group), (25±3), (50±3), (75±3) mg·kg^-1 respectively. The respiratory chambers controlled the environment with full-automation. The entire experimental period was 21 days: the starter phase was 21-31 d, and the later phase was 32-42 d. Broilers were provided ad libitum access to feed and water in nets. The antioxidant capacity of blood and muscle, and meat quality were determined at 32 d and 42 d of age.【Result】(1) Antioxidant capacity: At 21-31 d of age, with ammonia concentration increasing, the activity of SOD showed a quadratic increase (P＜0.05), and the activity of T-AOC(total antioxidation capacity) and the content of MDA (malondialdehyde) showed a linear increase (P＜0.05) in serum. The activity of CAT (catalase) and the content of MDA in the breast muscle showed a linear increase trend (0.05＜P＜0.10), and the activity of T-AOC showed a linear decrease (P＜0.05) in breast muscle. The activity of T-AOC showed a linear increase trend (0.05＜P＜0.10) in thigh muscle of broilers, and the activity of SOD (superoxide dismutase) of broilers in 75 mg·kg^-1group was higher than in others (P＜0.05). The activity of CAT in liver showed a linear decrease trend (0.05＜P＜0.10), the content of MDA showed a linear increase (P＜0.05). At 32-42 d of age, with ammonia concentration increasing, the activity of GSH-Px (glutathione peroxidase) showed a quadratic decrease (P＜0.05), and the activity of CAT showed a linear decrease in serum of broiler (P＜0.05). The activity of SOD in serum in the 50 mg·kg^-1 group was lower than that in the control (P＜0.05). The activity of CAT showed a linear increase (P＜0.05), and activity of T-AOC showed a quadratic decrease in breast muscle (P＜0.05). The activity of GSH-Px showed a quadratic increase (P＜0.05) in thigh muscle of broilers. The activity of SOD in thigh muscle of broilers exposed to 75 mg·kg^-1 was significantly higher than other groups (P＜0.05). With ammonia concentration increasing, the activity of T-AOC in the liver showed a quadratic increase trend (0.05＜P＜0.10). The activity of GSH-Px showed a linear decrease in liver (P＜0.05). (2) Meat quality: At 21-31 d of age, with ammonia concentration increasing, the shear force and the values of b* showed a linear decrease (P＜0.05), on the contrary, the values of L* showed a linear increase in breast muscle (P＜0.05), but the values of L* in the thigh muscle showed a linear decrease (P＜0.05), and the values of a* and pH 24 h showed a quadratic increase (P＜0.05). The values of b* showed a linear decrease trend (0.05＜P＜0.10). At 32-42 d of age, with ammonia concentration increasing, the shear force and the values of a* showed a linear decrease (P＜0.05), but the values of b* and pH 24 h showed a linear increase in breast muscle (P＜0.05). The values of L* in breast muscle of broilers exposed to 50 mg·kg^-1 were significantly higher than the other groups. The values of pH 45 min in thigh muscle showed a quadratic increase trend (0.05＜P＜0.10).【Conclusion】The result suggested that, ammonia in broiler houses adversely affects on antioxidant capacity and meat quality. Moreover, the adverse effects of ammonia could be worse with increased concentration and exposure period.

Development of a Microsphere-Based Fluorescence Immunochromatographic Assay for Detection of Kanamycin

XU Fei, ZHOU Jie, WU Chao, WANG Jian-fen, DING Shuang-yang, LI Xiu-bo

Scientia Agricultura Sinica. 2015, 48(21):  4358-4365.  doi:10.3864/j.issn.0578-1752.2015.21.015

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【Objective】 Kanamycin is a common drug that is used for the treatment of bacterial diseases in dairy cattle, and its residue in milk can not be ignored. Fluorescent microsphere, as a new type of fluorescence tracer, has presented a unique advantage towards the detection of small molecules in recent years. This study introduces a fluorescent microsphere immunochromatographic quantitative method for detecting kanamycin in milk, which provides a rapid and effective screening means for monitoring kanamycin residues. 【Method】 Two coating antigens of KANA (KANA-GA-OVA₁, KANA-I2-OVA₂) were developed by glutaraldehyde method and NaIO₄ method. After purification of anti-KANA monoclonal antibody by protein G immune affinity column, the antibody was coupled with fluorescent microspheres using EDC for the preparation of antibody-fluorescent microsphere conjugates (FM-mAbs). The immunochromatographic strip test was conducted as follows: NC membrane was coated with kanamycin antigen as a reaction carrier, (FM-mAbs) incubated with the sample, and allowed to migrate from below. The drug in-sample and coating antigen compete with each other for the limited FM-mAbs, and the amount of drug was evaluated by detection of the fluorescence intensity of FM-mAbs combined with coating antigen. 【Result】 It shows that the fluorescence intensity of test line gradually weakened with increasing concentrations of KANA on the sensitivity test, when drug standard solutions were added into samples at 0, 6.25, 12.5, 25, 50, 100 and 200 µg?L^-1. The F/F₀ ratio was selected to express the competitive inhibition, where F₀ and F are the fluorescence intensities respectively obtained from binding at zero and certain concentrations of the KANA standard. Standard curves were obtained by plotting F/F₀ against the analyte concentration and fitted to a four-parameter logistic equation using Originpro 8.0 software. The dynamic range IC₂₀-IC₈₀ was calculated as 9.5-100 µg?L^-1, and the IC₅₀ and LOD (IC₁₀) were 24.8 and 5.0 μg?L^-1, respectively. When the blank samples were spiked at 25, 50 and 100 µg?L^-1, the mean recoveries ranged from 78.4%-92.7%, with intra-assay coefficient of variations ranging from 10.8%-12.4%. The specificity of the method was evaluated by determining the cross-reactivity using other common aminoglycoside drugs. Negligible cross-reactivity (＜1%) was found for these other aminoglycosides. 【Conclusion】 The method is a rapid, sensitive, simple, effective, specific and suitable for promotion and application.

Dynamic Analysis of Expression Level of miR-451 and MIF Gene in Different Developmental Stages of Adult Hyalomma asiaticum

LUO Jin, YUAN Xiao-song, HAO Jia-wei, TIAN Zhan-cheng, XIE Jun-ren, CHEN Ze, REN Qiao-yun, YIN Hong, LUO Jian-xun, LIU Guang-yuan

Scientia Agricultura Sinica. 2015, 48(21):  4366-4373.  doi:10.3864/j.issn.0578-1752.2015.21.016

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【Objective】The microRNAs (miRNAs) are a class of small single-stranded noncoding RNAs (18—25 nt) in both unicellular and multicellular organisms. Previous studies showed that miRNAs play pivotal roles in regulating diverse developmental processes by targeting mRNAs for translational repression, development, or transcription. To dissect the interaction between miR-451 and target gene (Macrophage Migration Inhibitory Factor, MIF), Hyalomma asiaticum ticks were analyzed. 【Method】The primers were designed for stem-loop and PCR based on the mature sequence of miR-451（AAA CCG UUA CCA UUA CUG AGU UU）from High-through sequence results of Hyalomma asiaticum. The mature sequence of miR-451 was amplified by RT-PCR and analyzed miR-451 sequences from different species. The dsRNA of miR-451 was obtained by gene synthesis and it was injected into unfed-adult H. asiaticum. Referencing the MIF gene of Amblyomma americanum in GenBank (Accession number: AF289543.2) for specific primers of qPCR to H. asiaticum, and the β-actin gene as an internal reference gene. After injecting dsRNA of miR-451, the MIF and miR-451 expression level of various developmental stages of unfed-adult ticks were detected by SYBR Green real-time RT-PCR. 【Result】The miR-451 PCR product was consistent with the prediction that it is 72 nt by agarose gel electrophoresis. The result showed miR-451 has a high conservatism in different species by multiple sequence alignment analysis, only Monodelphis domestica has a base mutations (A→U) at 19 sites. Our results showed that the miR-451up-regulation at 0-30h and 6h had highest expression level with a copy number of 1.0×10⁸. Subsequently, the expression level gradually reduced, such that they had same level at 30 h with PBS control. The expression had an up-regulation until 48-60 h again. After the miR-451 was restrained the MIF was up-regulated at 30 h, and 42 h peaks (with copy levels of 9.0×10³), since the expression is restrained, and the level is a same with PBS control at 48. At 84 h the expression level of MIF was up-regulated and the peak was at 90 h. All the expression levels showed a normal distribution. 【Conclusion】 In this paper, the miR-451 sequence was amplified by RT-PCR, and we showed that miR-451 had high conservatism across different species. The conservatism of miR-451determined the target gene specificity. These results suggested that miR-451 plays an important biological function in animal cells and is an important regulatory factors for MIF function. As a target gene of miR-451 functions of MIF was regulated by miR-451using the RNAi, and they are reverse regulated. When the expression level of miR-451 is up-regulated, MIF expression were significantly lowered. The study is the first to confirm that expression of miR-451 is a common phenomenon in developmental stages of H. asiaticum ticks and that miR-451 has a negative regulatory effect on MIF. This study provides a reference for miR-451participation in immune response of ticks and the interaction mechanism between miR-451and target genes. It also confirms that the miRNA regulatory behavior has specificity and offers scheduling for gene function.

RESEARCH NOTES

Agronomic and Physiological Characterization of the Wide Adaptable Wheat Cultivar Zhongmai 175 Under Two Different Irrigation Conditions

LI Xing-mao, NI Sheng-li

Scientia Agricultura Sinica. 2015, 48(21):  4374-4380.  doi:10.3864/j.issn.0578-1752.2015.21.017

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【Objective】This study aims to differentiate the agronomic and physiologic characteristics among three winter wheat cultivars under water-limited and full irrigated conditions, and to identify the traits closely associated with the adaptability of Zhongmai 175, which will provide the reference in a breeding program for improving wheat adaptability.【Method】Three cultivars were grown in the 2011-2012 and 2012-2013 cropping seasons at the CAAS, Beijing Experimental Station, and at Gaoyi in Hebei province. Randomized complete blocks were used to incorporate water in two levels with two replications. Heading date, plant height, grain yield and components, ground cover after green recover stages (GC), normalized difference vegetation index before heading (NDVIv) and at mid-filling (NDVIg), SPAD before heading (SPADv) and at mid-filling（SPADg), leaf senescence score at mid-filling (LSS), leaf area index before heading (LAIv) and at mid-filling (LAIg), leaf senescence rate (LSR), and leaf rolling score (LRS) were measured. The differences of traits were analyzed among three varieties. 【Result】All traits except kernel number per spike, LAIg, and SPADv showed significant genetic variation. The values of all traits under full irrigation were increased more than those under limited irrigation. Grain yield, spike number per square meter, thousand kernel weight, plant height, and heading date of Zhongmai 175 were increased 1 072.6 kg·hm^-2, 104, 0.9 g, 7.2 cm, and 3 days, while those of Jingdong 8 were 1 274 kg·hm^-2, 15, 1.8 g, 9 cm, and 2 days, and Aikang 58 were 991.7 kg·hm-2, 47, 1.7 g, 6.2 cm, and 3 days. Grain yield and spike number per square meter of Zhongmai 175 were 647.3-1 505.8 kg·hm^-2 and 38-68.8 higher than other cultivars. Plant height, GC, NDVIv, NDVIg, SPADg, LSR, and LRS of Zhongmai 175 were 75.8 cm, 13.2%, 0.64, 0.55, 56.5, 14.10, and 4.0, respectively. The water sensitive index of Zhongmai 175 was 0.86, 0.64, 0.94, 0.61, and 0.57 for yield, thousand kernel weight, plant height, SPADg, and LAIv, respectively. 【Conclusion】 Zhongmai 175 showed the highest yield and spike number per unit area, early maturity, and semi-dwarf in limited irrigation and full irrigation areas. It was characterized by higher ground cover after the green recover stages, but lower normalized difference vegetation index in the heading and mid-filling stages, higher SPADg, and higher leaf rolling score. The grain yield, thousand kernel weight, plant height, SPADg, and LAIv were insensitive to water, but the spike number and kernel number of Zhongmai 175 were sensitive.

Effect of Organic and Inorganic Fertilizer on the Emission of CO₂ and N₂O from the Summer Maize Field in the North China Plain

LI Yan-qing, TANG Ji-wei, CHE Sheng-guo, WEN Yan-chen, SUN Wen-yan, ZHAO Bing-qiang

Scientia Agricultura Sinica. 2015, 48(21):  4381-4389.  doi:10.3864/j.issn.0578-1752.2015.21.018

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【Objective】 Under equal N rates, chemical fertilizer was compared with organic fertilizer in terms of the emission of GHG (CO₂ and N₂O) and GWP to properly understand the contribution of chemical fertilizer and organic fertilizer in greenhouse gas (GHG) emission in the field. The present research will be helpful in establishing GHG emission reduction measures.【Method】In a 8 year field experiment, CO₂ and N₂O emission flux from the Fluvo-aquic soil of summer maize were measured for 5 months (from June to October in 2014) by using a static chamber-gas chromatograph technique, and GHG effect and total season emission were also estimated. 【Result】Both the chemical fertilizer treatments and the organic fertilizer treatments had similar N₂O emission curves. A temporary peak of N₂O emission flux occurred after fertilization, and then the curves became stable. Under equal N rates, the N₂O flux of chemical fertilizer treatments were significantly higher than that of organic fertilization treatments, but organic fertilization treatments were higher than chemical fertilizer treatments in a stable phase after the peaks. Chemical fertilizer had little influence on CO₂ emission curves. Continuous peaks of CO₂ emission flux occurred after application of organic fertilization. Both the organic fertilizer and chemical fertilizer can increase the emission of N₂O. The emission of N₂O increased with the increase of nitrogen application. Under equal N rates, total N₂O emissions of chemical fertilizer treatments were significantly higher than organic fertilizer treatments. Application of organic fertilizer significantly increased CO₂ emissions, but the effect of chemical fertilizer on CO₂ emissions were not significant. When the N fertilizer rate was 240 kg·hm^-2 , both the chemical fertilizer treatments and organic fertilizer treatments had high level yields and a minimum GHGI, that were 0.27, 0.63 kg·hm^-2. Treatments with a higher N rates than 240 kg·hm^-2 had large GHGI values.【Conclusion】Application of more chemical or organic fertilizer would increase the emission of more greenhouse gas. Due to the carbon sequestration effect of the organic fertilizer, organic fertilizer treatments had smaller GHGI values than chemical fertilizer treatments, the suitable rate of organic fertilizer was an important solution to sequestrate carbon and reduced GHG emission.

iTRAQ-Based Proteomic Analysis for Identification of Candidate Proteins Underlying Beak Deformity in Chickens

LIU Nian, SUN Yan-yan, BAI Hao, HUA Deng-ke, XUE Fu-guang, LIU Ran-ran, LI Dong-li, WEN Jie, CHEN Ji-lan

Scientia Agricultura Sinica. 2015, 48(21):  4390-4396.  doi:10.3864/j.issn.0578-1752.2015.21.019

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【Objective】Beak deformity has been reported in some indigenous chickens like Beijing-You chickens and Qingyuan partridge chickens with deformed beaks have problems with eating and drinking resulting in obvious reductions of growth and production performances. Observations on pedigreed individuals with deformed beaks shed light on the heritability of this trait. The aim of this study was to identify the candidate proteins related to beak deformity using isobaric tags for relative and absolute quantitation (iTRAQ) technology and bioinformatic analysis. The results can build the basis for further understanding the mechanism of beak deformity in chickens. 【Method】Three chickens with deformed beaks (W1, W2 and W3) and their sibs (Z1, Z2 and Z3) of 120 days of age were used in this study. Beak sample proteins of these chickens were collected by using tissue total protein lysis buffer. Six iTRAQs were used to label samples of three contrast groups (W1 vs Z1, W2 vs Z2, and W3 vs Z3). After separation by chromatography and analysis of liquid chromatography tandem mass spectrometry, differentially expressed proteins were identified and quantitatively analyzed by Mascot 2.3.02 software. Comparing deformed samples with the normal samples, we chose some differentially expressed proteins which consisted of 2 or more peptides, P＜0.05, and the ratio of which was more than 1.2 (up-regulated) or less than 0.83 (down-regulated). 【Result】 In total, 3 372 proteins were detected from three contrast groups using iTRAQ and 1 869 of them were Jungle fowl proteins. Of these, 159 were differentially expressed proteins, 70 were up-regulated and 89 were down-regulated. The most up-regulated proteins included LPL, MLC-2, CO9A1, MATN3, and HSP90B1, while the most down-regulated proteins were MBP, RLA1, PRVM, and HAPLN1. Protein CO9A1, MATN3, and HAPLN1 were associated with cartilage synthesis and ossification. PRVM is an intracellular calcium binding protein involved in regulating the Ca²⁺ ion signaling pathway. LPL is a multifunctional enzyme and mainly plays a role in lipid metabolism and transport, which involves regulating PPAR signaling pathways. 【Conclusion】 Proteins CO9A1, MATN3, HAPLN1, PRVM, and LPL were proposed as candidate proteins for beak deformity. The discovery and identification of these proteins can offer a valuable insights into basis of protein level of beak deformity in chickens.