Scientia Agricultura Sinica

Genetic Analysis and Mapping of Novel Cleistogamy Gene, cl7(t) in Rice

NI Da-Hu, YANG Ya-Chun, SONG Feng-Shun, NI Jin-Long, LI Li, YANG Jian-Bo

Scientia Agricultura Sinica. 2012, 45(13): 2561-2567. doi:10.3864/j.issn.0578-1752.2012.13.001

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【Objective】 Identification, genetic analysis and mapping of novel cleistogamy genes in rice would play an important role in understanding the molecular genetic mechanisms of floral development. At the same time, the cleistogamy is an ideal strategy to prevent pollen explosion in transgenic rice. 【Method】 A rice cleistogamy mutant 8m30 with japonica cultivar H02 background was isolated in M2 population treated with 0.5% EMS (Ethane Methyl Sulfonare). Phenotypic study, genetic analysis and gene mapping by map-based cloning were conducted with the population from F1 of 8m30 and Guanghui102 and its self-cross progenies.【Result】The mutant with lower plant height, compatible plant style and closed paleas at flowering stage. Genetic analysis indicated that the mutant was controlled by a single recessive nuclear gene. The cleistogamy locus was mapped on the long arm of chromosome 7 between the microsatellite markers RM21964 and RM234 with 0.1 cM and 0.3 cM genetic distance, about 160 kb of the physical distance and cosegregated with RM21971, which suggests the mutant locus be novel allele and is tentatively named cl7(t) (Cleistogamy 7 (t)). 【Conclusion】 The 8m30 mutant is controlled by a recessive nuclear gene, which is located on chromosome7 between RM21964 and RM234 with physical distance of 160 kb.

Genetic Analysis and QTL Mapping of Pod-Seed Traits in Soybean Under Different Environments

LIANG Hui-Zhen, YU Yong-Liang, YANG Hong-Qi, ZHANG Hai-Yang, DONG Wei, LI Cai-Yun, GONG Peng-Tao, LIU Xue-Yi, FANG Xuan-Jun

Scientia Agricultura Sinica. 2012, 45(13): 2568-2579. doi:10.3864/j.issn.0578-1752.2012.13.002

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【Objective】 Soybean [Glycine mar (L.) Merr.] pod-seed trait is one of the most important traits related with yields. In this paper, the relationship between soybean pod-seed traits and yields was studied, the QTLs controlling soybean pod-seed traits were mapped, and the soybean pod-seed trait molecular genetic mechanism was revealed. 【Method】Based on three years and two environments data, a recombinant line population of 474 F2:8:11 lines from a cross of Jindou 23 and ZDD2315 was used for analyzing and mapping QTLs of soybean pod-seed related traits. 【Result】The results of the molecular genetic analysis indicated that the field traits have a significant positive correlation or extreme significant positive correlation with 100-grain-weight, grain length, single grain weight, two-grain pod and three-grain pod. According to three years and two environments data, the authors mapped six QTLs of field traits related, four QTLs of 100-grain-weight, ten QTLs of single line grain weight, two QTLs of grain width, five QTLs of grain length, seven QTLs of one-grain pod, five two-grain pod, seven QTLs of three-grain pod and five QTLs of four-grain pod. 【Conclusion】The identification of consistent QTLs under different environments and in different years will be used as candidate chromosome regions, which will help to improve the efficiency of selection for pod-seed related traits in soybean through molecular marker-assisted selection.

Association Mapping of Sesame (Sesamum indicum L.) Resistance to Macrophomina phaseolina and Identification of Resistant Accessions

ZHANG Yan-Xin, WANG Lin-Hai, LI Dong-Hua, WEI Wen-Liang, GAO Yuan, ZHANG Xiu-Rong

Scientia Agricultura Sinica. 2012, 45(13): 2580-2591. doi:10.3864/j.issn.0578-1752.2012.13.003

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【Objective】Developing molecular markers related with resistance to Macrophomina phaseolina from natural population, identifying elite resistant accessions. 【Method】In this study, 216 sesame core collections were scanned by 79 pairs of EST-SSR, SRAP and AFLP primers, accurate identification of their resistance to Macrophomina phaseolina was carried out from 2009 to 2011, and based on these data the genes related with resistance to M. phaseolina were located by association analysis. 【Result】A total of 608 polymorphic bands were amplified, population structure analysis indicated that it was consisted of 2 subgroups, 43 markers significantly associated with M. phaseolina disease index simultaneously in 2 or 3 years were detected by association analysis based on GLM(Q) and MLM(Q+K) model, explained 1.82% to 9.46% of phenotypic variation, 3 associated markers were detected by both of 2 models, and marker M7E6-1 was presented simultaneously in 3 years. Ten accessions with resistance to M. phaseolina were identified, the coincident rate of 43 associated markers in the 10 accessions was 60% to 100%, with an average of 92.09%. 【Conclusion】 The tested population was consisted of 2 subgroups. There was no relationship between division of subgroup and geographical origin of the material, 43 markers were detected significantly and stably associated with M. phaseolina disease index. There was a diverse variation among the resistance to M. phaseolina of the tested population, and 10 accessions with stable resistance to M. phaseolina were identified.

Effect of Sowing Date on Characteristics of Photosynthesis and Matter Production of Direct Seeding Rice

HUO Zhong-Yang, YAO Yi, ZHANG Hong-Cheng, XIA Yan, NI Xiao-Cheng, DAI Qi-Gen, XU Ke, WEI Hai-Yan

Scientia Agricultura Sinica. 2012, 45(13): 2592-2606. doi:10.3864/j.issn.0578-1752.2012.13.004

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【Objective】The objective of the study is to elucidate the effect of sowing date on characteristics of photosynthesis and matter production of direct seeding rice.【Method】Taking four representative rice cultivars of three types of early-maturing late japonica rice, late-maturing medium japonica rice and medium-maturing medium japonica rice (including japonica conventional rice and japonica hybrid rice) as experimental materials, the dry matter accumulation, distribution, translocation and leaf area, photosynthetic potential, crop growth rate and net assimilation rate of direct seeding rice were studied.【Result】Along with the delayed sowing date, dry matter weight of single stem and population at jointing, booting, heading, waxy stages and maturity both showed a apparent downward trend, and the difference was increasing with the growing process. Along with the delayed sowing date, the proportion of dry matter accumulation from sowing to jointing and from jointing to heading increased, but from heading to maturity decreased, while the dry matter accumulation of every stage reduced significantly or very significantly. With the growing process, leaf weight per stem and sheath weight per stem decreased, stem weight per stem firstly increased and then decreased, and panicle weight per stem increased. Along with the delayed sowing date, leaf weight per stem and sheath weight per stem decreased significantly or very significantly at booting and heading, little difference at waxy stage and increased a little at maturity, stem weight per stem and panicle weight per stem decreased significantly or very significantly at all stages. With the growing process, the proportion of leaf, stem, sheath and panicle in the later growth period had the same changes as the dry matter weight. Along with the delayed sowing date, the ratio of leaf and sheath decreased a little at booting, little difference at heading and increased significantly or very significantly at waxy stage and maturity, the ratio of stem decreased a little at booting and heading, increased a little at waxy stage and decreased at maturity, the ratio of panicle increased a little at booting, little difference at heading and decreased significantly or very significantly at waxy stage and maturity. Along with the delayed sowing date, the exportation, export rate and transformation rate of leaf, stem and sheath decreased significantly or very significantly. Along with the delayed sowing date, the leaf area index had a little difference at jointing, decreased significantly or very significantly at booting, heading and waxy stages, and decreased a little at maturity, the photosynthetic potential from sowing to jointing, jointing to heading and the whole growth period decreased significantly or very significantly, and decreased from heading to maturity, but the difference was little not significant. Along with the delayed sowing date, the crop growth rate from sowing to jointing increased significantly, little difference from jointing to heading and decreased significantly from heading to maturity, the net assimilation rate from sowing to jointing and from jointing to heading increased obviously, but decreased obviously from heading to maturity.【Conclusion】Sowing date had a great effect on characteristics of photosynthesis and matter production of direct seeding rice. Compared to sowing late, sowing early has the advantages that the dry matter accumulation is appropriate in the early stage and significantly higher in the middle and late stages, so the total dry matter accumulation is significantly higher and its distribution is reasonable, the export and transformation rate is high, and the photosynthetic production capacity is great after heading.

Spatial Distribution of Leaf and Boll in Relation to Canopy Photosynthesis of Super High-Yielding Cotton in Xinjiang

FENG Guo-Yi, LUO Hong-Hai, YAO Yan-Di, YANG Mei-Sen, DU Ming-Wei, ZHANG Ya-Li, ZHANG Wang-Feng

Scientia Agricultura Sinica. 2012, 45(13): 2607-2617. doi:10.3864/j.issn.0578-1752.2012.13.005

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【Objective】To explore the changes of leaf area distribution, internodes length and leaf inclination angle, the influence of leaf spatial distribution on canopy structure and its correlation with canopy photosynthesis was discussed, and the formation mechanism of super-high-yielding cotton was studied as well.【Method】The methods of definite cultivating super-high-yielding cotton (lint above 4 000 kg•hm-2) were adopted in this paper. The spatial distribution of leaf area index, internode length, and leaf inclination angle were measured at different growth stages and the effects of canopy structural changes on canopy apparent photosynthesis in relation to bolls spatial distribution were analyzed.【Result】The properties of super-high-yielding cotton (lint above 4 000 kg•hm-2) canopy were that the plant height was 72.3-87.7 cm before boll opening stage and the average internodes length was 7.15-7.20 cm. Moreover, the internode length in the upper and middle layers of plant was longer. The leaf inclination angle in upper, middle and lower layers of plant was 48.8-53.8, 41.0-49.3 and 30.1-40.1, respectively. The proportion of leaf area index between upper, middle and lower layer of plant was about 1:1:1 from flowering stage to later full bolling stage. At boll opening stage, the leaf area index and canopy photosynthetic rate in upper layer of plant remained 0.95-1.76 and 8.1-13.2 μmol•m-2•s-1, respectively. The proportion of canopy photosynthetic rate in upper layer to whole layers was 45.9%-59.8%. The proportion of the boll number between upper, middle and lower layers of plant was 1.8﹕1.2﹕1, and there were more boll numbers and higher sink potentials in the upper layer of plant.【Conclusion】The physiological basis of super high-yielding cotton formation was that longer internode length and larger leaf inclination angle in upper and middle layers and higher proportion leaf area index in middle and lower layers, the leaf canopy photosynthetic rate was higher and equidistribution in vertical direction from flowering to later full bolling stage. In addition, the leaf area index and canopy photosynthetic rate declined slowly at the boll opening stage. Taken together, the spatial distribution anastomosis of leaf canopy photosynthetic rate and boll, and consequently, the coordinate development of leaf area-boll were proposed.

Identification of Pathogens Causing Common Bacterial Blight on Common Bean (Phaseolus vulgaris L.)

CHEN Hong-Yu, XU Xin-Xin, DUAN Can-Xing, WANG Shu-Min, ZHU Zhen-Dong

Scientia Agricultura Sinica. 2012, 45(13): 2618-2627. doi:10.3864/j.issn.0578-1752.2012.13.006

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【Objective】The objective of this study is to identify the pathogens that cause common bacterial blight on common bean in north China.【Method】 The causal agents were identified by colony characterization, detection using Xanthomonas campestris pv. phaseoli diagonsis kit (ADGEN EXPRESSTM), pathogenicity test, sequencing 16 rDNA and 16S-23S rDNA intergenic spacer (ITS) regions, PCR detection using specific primers for X. axonopodis pv. phaseoli and X. fuscans subsp. fuscans, fatty acid methyl esters (FAME) analysis and biochemical characterization tests.【Result】Bacterial isolates of Xanthmonads-like were isolated from all diseased plant and seed samples. Twenty-five representative isolates were selected for pathogenicity test, and 24 tested isolates casused typical symptoms of common bacterial blight on the inoculated plants of common bean cutivar ‘Yingguohong’. Based on pathenogenicity test, and combined with the results of diagonsis kit and specific PCR detection, biochemical and molecular characterization, 7 isoaltes were identifed as X. axonopodis pv. phaseoli, and 17 isolates were identified as X. fuscans subsp. fuscans.【Conclusion】 The common bacterial blight on common bean in north China was incited by X. axonopodis pv. phaseoli or/and X. fuscans subsp. fuscans. It was the first report of X. fuscans subsp. fuscans in China.

Effects of Cuminic Acid on the Growth and Development of Phytophthora capsici Leonian

FENG Jun-Tao, HAN Li-Rong, FAN Rui-Juan, CHEN Cong-Zhen, ZHANG Xing

Scientia Agricultura Sinica. 2012, 45(13): 2628-2635. doi:10.3864/j.issn.0578-1752.2012.13.007

Abstract ( 714 )

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【Objective】 The objective of this study is to explore the activity of cuminic acid against the growth of Phytophthora capsici.【Method】Agar plate tests, pendant-drop method and pot experiments were conducted to test the inhibitory effect on P. capsici in vitro and in vivo of cuminic acid. The ultrastructure of P. capsici treated with cuminic acid was observed through scanning electron microscope (SEM) and transmission electron microscope (TEM). And the distribution of the main components, β-1,3-glucan content and β-1,3-glucanase activity of the P. capsici cell wall exposed to cuminic acid were studied using Calcofluor white staining and phenol sulfuric acid method. 【Result】 Cuminic acid displayed strong antifungal activity to P. capsici in vitro and in vivo, and the germination of zoospores was strongly inhibited. The observations with SEM revealed that the excessive branching mycelium occurred, the surface of hyphae was tumorous protuberance, irregular swelling and intensive growing point. The sporangiums were invaginated, and some of them even collapsed and formed shells. The observations with TEM showed that the hyphal cell wall became thickening irregularly and the color was deepened. The mitochondria swelled, crest in some of them had disappeared, some small organelles such as endoplasmic reticulum and ribosomal were losing their structures. Moreover, the polar deposition of newly synthesized cell wall components in germinating cystospores was disturbed and the β-1,3-glucan content and β-1, 3-glucanase activity decreased significantly. 【Conclusion】 Cuminic acid significantly inhibited the growth and development of P. capsici, so it is worthy of further study.

Degradation Dynamics and Residues Analysis of Chlorantraniliprole in Cabbage and Soil by QuEChERS-High Performance Liquid Chromatography-Tandem Mass Spectrometry

CHEN Xiao-Jun, WANG Meng, FAN Shu-Qin, CUI Hai-Rong, YANG Yi-Zhong

Scientia Agricultura Sinica. 2012, 45(13): 2636-2647. doi:10.3864/j.issn.0578-1752.2012.13.008

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【Objective】 The objective of this study is to research degradation dynamics of chlorantraniliprole in the vegetable and soil, and to formulate a scientific basis for the reasonable spray volume and the safe interval period on controlling vegetable pests.【Method】Degradation dynamics and terminal residue of chlorantraniliprole of 20% chlorantraniliprole SC in cabbage and soil were investigated by QuEChERS-high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS).【Result】 The recoveries of chlorantraniliprole from the cabbage and the soil were 81.25%-92.05% and 82.92%-93.38%, respectively. The residues in the cabbage and soil were proved to chlorantraniliprole through qualitative analysis by HPLC-MS/MS. Degradation dynamic equations of chlorantraniliprole in cabbage and soil fitted the first-order reaction dynamic equations. The half-lives of degradation of chlorantraniliprole in cabbage and soil were 7.66 and 6.86 d, respectively. The final residues of chlorantraniliprole were not detected in cabbage and 0.0071 mg•kg-1 in soil, respectively, when 20% chlorantraniliprole SC was applied at 0.045 g•m-2. The final residues of chlorantraniliprole were 0.0063 mg•kg-1 in cabbage and 0.1004 mg•kg-1 in soil, respectively, when 20% chlorantraniliprole SC was applied at 0.090 g•m-2.【Conclusion】20% chlorantraniliprole SC was applied at 0.045 and 0.090 g•m-2 could be considered safe in the cabbage field.

Effects of No-Tillage and Rice-Seedling Casting with Rice Straw Returning on Content of Nitrogen, Phosphorus and Potassium of Soil Profiles

HUANG Jing, GU Ming-Hua, XU Shi-Hong, YANG Wei-Fang, JIANG Li-Geng

Scientia Agricultura Sinica. 2012, 45(13): 2648-2657. doi:10.3864/j.issn.0578-1752.2012.13.009

Abstract ( 810 )

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【Objective】 The objective of this study was to understand the effects of no-tillage and rice-seedling casting with rice straw returning on nitrogen, phosphorus and potassium of paddy soil profile.【Method】Contents of nitrogen, phosphorus and potassium of paddy soil profile soil of conventional tillage of rice-seedling casting and no-tillage of rice-seedling casting experiments that lasted two and seven years were investigated. 【Result】Contents of total nitrogen, total phosphorus, alkaline-hydrolysable nitrogen and rapidly available phosphorus in 0-4(5) cm soil layer of no-tillage treatments were higher than those of conventional tillage. Contents of total potassium in 0-4(5) cm soil layer of no-tillage with rice straw returning treatment were significantly higher than those of common conventional tillage. Contents of rapidly available potassium in 0-4(5) cm soil layer of no-tillage with rice straw returning treatment were lower than that of conventional tillage with rice straw returning, but significantly higher than that of common conventional tillage. Contents of total nitrogen, total phosphorus, total potassium, alkaline-hydrolysable nitrogen and rapidly available phosphorus in 5-20 cm soil layer of no-tillage treatments were lower than those of conventional tillage. Content of rapidly available potassium in Aa layer (cultivation layer) of common no-tillage treatment was lower than those of conventional tillage. Contents of alkaline-hydrolysable nitrogen and rapidly available potassium in Ap layer (plow pan), W layer (waterlogged layer) and C layer (parent material layer) were higher than those of conventional tillage. 【Conclusion】The effects of no-tillage of rice-seedling casting with of rice straw returning on coordinate soil nitrogen, phosphorus and potassium are superior to that of common no-tillage of rice-seedling casting, and no-tillage of rice-seedling casting with rice straw returning is advantageous to the improvement and development of soil fertility. No-tillage of rice-seedling casting with rice straw returning is a development direction of paddy field no-tillage.

Effects of Earthworms Collected from South China on Soil Enzyme Activities and Microbial Characteristics

ZHANG Chi, CHEN Xu-Fei, ZHOU Bo, LI Jian-Long, YANG Cheng-Fang, DAI Jun

Scientia Agricultura Sinica. 2012, 45(13): 2658-2667. doi:10.3864/j.issn.0578-1752.2012.13.010

Abstract ( 823 )

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【Objective】Effects of two earthworm species on soil microbial characteristics and enzyme activities were studied in paddy and vegetable soils in South China. 【Method】Two earthworm species (Amynthas robustus and Amynthas corticis) were cultured in paddy and vegetable soils in the lab. Parameters such as soil catalase, urease, invertase, acid and alkaline phosphatase, soil basal respiration, microbe biomass carbon, microbial metabolic quotient were analyzed to evaluate earthworm’s effects on soil biological quality in different soil types. 【Result】Results showed that A. corticis could significantly improve soil basal respiration, enhance catalase and ureased activities, and decrease acid phosphatase activities (P＜0.05) in paddy soil. In vegetable soil, both of earthworm species increase urease activities. A. robustus and A. corticis increased invertase and catalase activities (P＜0.05), respectively. Multivariate analysis showed that A. corticis and A. robustus improved soil biological properties in both of two kinds of soil, and especially more positive effects on soil biological quality were shown in A. corticis than A. robustus in vegetable soil. 【Conclusion】Earthworms improved soil biological quality in the study, so it could be considered as a biological resource for the application in sustainable soil management. Effects of earthworms on soil biological quality depend on their ecological categories and soil properties. It is recommended that more studies on the screening of earthworm species and organic matter additions should be performed in future according to the local conditions.

Comparison of Gibberellin Acid Content and the Genes Relatived to GA Biosynthesis Between ‘Changfu 2’ Apple (Malus domestica Borkh.) and Its Spur Sport

SONG Yang, ZHANG Yan-Min, LIU Jin, WANG Chuan-Zeng, LIU Mei-Yan, FENG Shou-Qian, CHEN Xue-Sen

Scientia Agricultura Sinica. 2012, 45(13): 2668-2675. doi:10.3864/j.issn.0578-1752.2012.13.011

Abstract ( 757 )

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【Objective】The relationship between internode length of shoots, endogenous hormones gibberellic acid (GA) and the structure and expression of genes related to GA synthesis in apple (Malus domestica Borkh.) is a basis for further exploration of the spur type development mechanism and breeding of cultivars of spur type apple. 【Method】 The shoots and leaves of ‘Changfu 2’ and its spur sport ‘Longfuduanzhi’ were used as experimental materials for measuring the content of GA and cloning the genes which involved in the GA synthesis pathway at the four time points days after flowering and researching the relationship between GA and the internode length of shoots at shoot development stage. 【Result】 The GA content was significantly different between the spur type apple and non-spur type apple on 80th day after flowering. The GA content was lower in spur type sport compared with in non-spur type apple varieties on 80th day after flowering. The sequences analysis showed that the cDNA sequences of GA20ox and KO, key genes involved in GA synthesis, were the same between ‘Longfuduanzhi’ and ‘Changfu 2’. The mutation, insertion and deletion were not appeared in GA20ox and KO. The real-time quantitative PCR analysis showed that the relative expression of GA20ox and KO were significantly different between ‘Longfuduanzhi’ and ‘Changfu 2’ on 20th and 80th day after flowering. The relative expression of GA20ox and KO was lower in spur type apple compared with non-spur type apple on 20th and 80th day after flowering. 【Conclusion】The research results showed that the difference of GA content and key genes expression of GA synthesis pathway were related to shoot internode length of spur sport apple. The lower GA content and genes involved in GA synthesis pathway were down-regulated expression which inhibited the normal development of spur sport apple shoots. This study has provided basic information for further exploration of the spur type development mechanism.

Construction of DNA Fingerprint Database Based on SSR Marker for Varieties(Lines) of Cucumis melo L.

SONG Hai-Bin, CUI Xi-Bo, MA Hong-Yan, ZHU Zi-Cheng, LUAN Fei-Shi

Scientia Agricultura Sinica. 2012, 45(13): 2676-2689. doi:10.3864/j.issn.0578-1752.2012.13.012

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【Objective】 DNA fingerprinting database of varieties of melon (Cucumis melo L.) was constructed and the identification method was obtained. 【Method】The simple sequence repeat (SSR) primers were selected using 20 representative melon varieties (lines) for establishing fingerprints of 105 different varieties (lines). 【Result】Out of the 1 219 pairs of SSR primers, 18 pairs were screened to generate polymorphic fingerprints among 105 varieties or inbred lines. The results showed that each pair of the primers could detect 4-14 polymorphic bands with an average of 9. The average polymorphism information content (PIC) was 0.68 with a range of 0.55-0.82. The genetic similarity coefficient of the 105 accessions ranged from 0.70 to 0.99. The fingerprint data of all the 18 pairs of core SSR primers were employed to establish the fingerprinting database to distinguish the 105 samples and every accession had its unique fingerprinting. 【Conclusion】 The results indicated that SSR marker is suitable to construction of DNA fingerprinting database of melon varieties and the database could provide reference for melon identification.

The cDNA Cloning and Analysis of Sequence Information and Quantitative Express of Chrysanthemum Rhythms Clock Output Gene CmGI (GIGANTEA)

SUN Xia, WANG Xiu-Feng, ZHENG Cheng-Shu, XING Shi-Yan, SHU Huai-Rui

Scientia Agricultura Sinica. 2012, 45(13): 2690-2703. doi:10.3864/j.issn.0578-1752.2012.13.013

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【Objective】The cDNA sequence of chrysanthemum rhythms clock output gene GIGANTEA was cloned, and the bioinformatics of the sequence and the relative quantitative expression of mRNA were analyzed.【Method】 Polymerase chain reaction (PCR) combined with 5′RACE, and 3′RACE technology were used to clone the full length cDNA of chrysanthemum rhythms clock output gene CmGI，analysis of sequence of nucleotides and code of protein was made by using the software of bioinformatics. Protein structure prediction of 3D modeling was made by using the online modeling software. The relative quantitative expression analysis of CmGI was conducted by real-time quantitative fluorescence PCR technology and 2-△△Ct method. 【Result】The cDNA sequence of GIGANTEA was cloned from chrysanthemum ‘Jniba’, the full-length cDNA was 3 461 bp, open reading frame (ORF ) was 3 453 bp, and encoded 1 150 amino acids. Sequence analysis showed that the genetic code of protein was homologous with plant rhythms clock output gene GIGANTEA, named CmGI gene. The sequence was submitted to GenBank, and the registration number is JQ043439. Sequence alignment displayed that it was a similarity of 76% and 75% with GIGANTEA of Vitis vinifera，Ricinus communis, respectively. The phylogenetic tree showed that chrysanthemum CmGI and Arabidopsis thaliana GIGANTEA are closest in molecular evolution distance, followed by Brassica rapa GIGANTEA. It was speculated that CmGI protein has six transmembrane spiral across a cell membrane many times. They are transcription factors, located in the nucleus and it is a non-secretory protein. They do not have a signal peptide. CmGI 3D structure modeling projections show that the protein core structure accords with the transcription factors and the function of the common DNA combining domain HTH and HLH. Fluorescent relative quantitative analysis shows that the expression patterns of chrysanthemum CmGI are circadian rhythms expression. At different flower bud differentiation stage, the CmGI gene in the leaf blade mRNA level is different, two peak values were appeared in the flower bud differentiation start-up and floret primordia middle differentiation periods. The tissue culture plantlets leaves and buds under long-day conditions, during alabastrum period are all trace expression and was in the order of leaves＞tongue shape flower＞tubular flowers during flowers blooming period. 【Conclusion】Rhythms clock output gene CmGI was cloned from chrysanthemum, further research on this gene will help exploration of photoperiod pathway of the flower of chrysanthemum molecular control mechanism and could be used as the target gene of molecular breeding of flowering phase.

Changes of Granule Structure and Enzyme Hydrolysability of Upper Flue-Cured Tobacco Leaves Starch During Bulk Flue-Curing

WANG Tao, HE Fan, TIAN Bin-Qiang, SONG Chao-Peng, SHI Long-Fei, XU Cheng-Long, ZHAN Jun, GONG Chang-Rong

Scientia Agricultura Sinica. 2012, 45(13): 2704-2710. doi:10.3864/j.issn.0578-1752.2012.13.014

Abstract ( 965 )

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【Objective】 In order to provide a theoretical basis for reducing the starch content and improving the quality of upper flue-cured tobacco leaves, changes of granule structure and characteristics of flue-cured tobacco leaves starch and the related enzymes were studied during bulk flue-curing.【Method】Changes of the activity of amylase, enzyme hydrolysability and granule structure of upper flue-cured tobacco leaves starch were studied during flue-curing using conventional physical and chemical analysis and SEM. 【Result】 The contents of the starch of tobacco leaves changed greatly at yellowing stage and the activity of amylase presented a double-peak curve during flue-curing. The SEM observation showed that most of the tobacco leaves starch were spheroidal and long cylindrical and some individual was irregular shape. The granule surface had obvious depression and layered structure. The layer structure appeared on the surface of large amounts of starch granules and the starch of long cylindrical was much reduced at yellowing stage. There was almost no existence of long cylindrical starch in flue-cured tobacco leaves. The average long-axis-diameter was 3.21 µm of fresh tobacco leaves starch. And the average long-axis-diameter of starch increased gradually during the flue-curing and it increased obviously at 38℃ and 42℃. The diameter of starch granule increased by more than 60% at the end of 42℃ and there was no remarkable difference after 47℃. Enzyme hydrolysability of flue-cured tobacco leaves starch increased with the proceeding of curing and there was a peak at 38℃ and then decreased. 【Conclusion】The yellowing stage was the critical period for the degradation and changes of granule structure and characteristics of flue-cured tobacco leaves starch during bulk flue-curing. It would have positive effects to improve the quality of upper flue-cured tobacco leaves by controlling the curing environments at yellowing stage.

Effect of Supplement of Soya Lecithine on Mucosal Anti-Oxidation and Heat Shock Protein 70 Content in Intrauterine Growth Retardation Piglets

WANG Yuan-Xiao, ZHANG Li-Li, ZHOU Gen-Lai, WANG Tian

Scientia Agricultura Sinica. 2012, 45(13): 2711-2717. doi:10.3864/j.issn.0578-1752.2012.13.015

Abstract ( 692 )

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【Objective】The effects of supplement of soya lecithine (SL) on intestinal growth, mucosal heat shock protein 70 (HSP70) expression and oxidation stress (OS) in intrauterine growth retardation (IUGR) piglets were studied.【Method】Twelve IUGR piglets and six piglets with normal birth weight were selected. All piglets were weaned on 7th d of age, and assigned equally to three groups: six NBW and six IUGR piglets were fed with control diet (IUGR), and six IUGR piglets were fed with diet supplemented with 1.5% SL (IUGR+SL) for 7 days.【Result】IUGR decreased (P＜0.05) the absolute weigh of jejunal segment and non-mucosa, reduced (P＜0.05) the mucosal capacity of total anti-oxidation (T-AOC), as well as the activity of GR and GPx, and elevated (P＜0.05) the content of MDA. The mucosal HSP70 contents detected by ELISA and Western Blot were increased (P＜0.05) by IUGR. After SL treatment, the absolute and relative weight of jejunum, jejunal mucosa and non-mucosa increased (P＜0.05) in IUGR piglets. Supplement of SL increased (P＜0.05) the mucosal T-AOC, GPx and SOD, as well as reduced (P＜0.05) MDA and HSP70 content in terms of ELISA. Inverse correlations were found between SL supplementation in IUGR piglets and HSP70 contents (P＜0.05).【Conclusion】SL plays an important role in recovering impaired jejunal growth, increased mucosal HSP70, and OS induced by IUGR.

Estimation and Regression Models of Methane Emissions from Sheep

ZHAO Yi-Guang, DIAO Qi-Yu, LIU Jie, JIANG Cheng-Gang, DENG Kai-Dong, TU Yan

Scientia Agricultura Sinica. 2012, 45(13): 2718-2727. doi:10.3864/j.issn.0578-1752.2012.13.016

Abstract ( 903 )

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【Objective】 This trial was designed to study the effects of dietary nutrients and their digestibility on rumen methane (CH4) emissions from sheep and develop regression models. 【Method】 Eight adult crossbreed rams (Dorper (♂) ×Small Tail Han sheep (♀)) with the body weight of (50.37±1.13) kg were fed 8 total mixed rations with different NDF content of 51.83%, 50.47%, 44.58%, 42.34%, 38.20%, 35.43%, 30.10% and 27.03%, respectively. The experiment design was 8×4 incomplete Latin square which lasted for 4 periods and each period lasted for 16 d. CH4 was measured by sable open-circuit respiration calorimetry system with 3 sheep once and digestive trials were conducted by total feces collection method during the last 8 d of each period. 【Result】 CH4 per kg digestive organic matter (L•kg-1 DOM) of 8 diets was 48.80, 46.98, 45.38, 41.34, 35.44, 34.66, 33.73 and 37.83, respectively. Methane regressed to NDF, ADF, nutrient intake and digestive nutrients were CH4E/DE (%)=0.14688DNDF/DOM (%)+5.47643 (R2=0.9084, P=0.0003), CH4 (L•kg-1 DOM)=0.14411DADF(g)+23.69940 (R2=0.8367, P=0.0015), CH4 (L•kg-1 DOM)=-0.22260OMI (g)+0.15234CPI (g)+0.06465NDFI (g)-0.60549EEI (g)+218.23715 (R2=0.9786, P=0.0077) and CH4 (L•kg-1 DOM)=0.09722DCP (g)+0.12079DNDF (g)-0.08355DADF (g)-1.71035DEE (g)+47.85414 (R2=0.9776, P=0.0083), respectively.【Conclusion】CH4 (L•kg-1 DOM), CH4E/GE and CH4E/DE were affected by the ratio of forage to concentrate (F﹕C) or dietary roughage content. In the linear regression models, NDF and ADF were more reliable than other nutrients to estimate methane emissions with higher precision. Compared with the linear regression models, the multiple regression models clearly provided a higher accuracy and a stronger correlation.

Polymorphism of Exon10 of FSHR Gene and Its Relationship with Litter Size in Xiaomeishan Pigs

WU Jing-Sheng, WANG Jin-Yu

Scientia Agricultura Sinica. 2012, 45(13): 2728-2736. doi:10.3864/j.issn.0578-1752.2012.13.017

Abstract ( 734 )

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【Objective】The effect of polymorphism of exon10 of FSHR gene on litter size in Xiaomeishan pigs was studied.【Method】The polymorphism of exon10 of FSHR gene was investigated by PCR-SSCP and direct sequencing methods, and the effect of exon10 of FSHR gene on litter size traits in Xiaomeishan pigs was analyzed using the least square analysis. Simultaneously the variance components were analyzed and selective reaction was predicted.【Result】 There was one novel polymorphic locus dected by primers P1 and P3, respectively, and the former made amino acid change, among Xiaomeishan, Fengjing and Large White pigs. One polymorphic locus detected by primer P6 was only in Large White pigs. Allele A at locus P1 and allele D at P3 locus were the dominant alleles in Xiaomeishan pigs. AA genotypic sow of Xiaomeishan pigs produced 1.95 TNB (total number born) and 1.66 NBA (number born alive) more than BB genotypic sow at locus P1 after the second parity, respectively (P＜0.01). And DD genotypic sow of Xiaomeishan pigs produced more TNB and NBA than CC genotypic sow at locus P3 after the second parity and in all parities, respectively (P＜0.01). The heredity of TNB and NBA in Xiaomeishan pigs was mostly influenced by the genetic additive effect.【Conclusion】Allele A at locus P1 and allele D at locus P3 in exon10 of FSHR gene had significant effect on litter sizes in Xiaomeishan pigs.

Breeding and Expression Stability of Human Lactoferrin Transgenic Goat

QIAO Na, XU Xu-Jun, LIU Si-Guo, WANG Xue-Bin, YU Hui-Qing, TONG Yong-Xing, CHEN Jian-Quan, CHENG Guo-Xiang

Scientia Agricultura Sinica. 2012, 45(13): 2737-2742. doi:10.3864/j.issn.0578-1752.2012.13.018

Abstract ( 673 )

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【Objective】 This study was conducted to analyze the effects of transgenosis on the reproduction of human lactoferrin (hLF) transgenic goat, and the genetic and expression stability of exogenous gene in each generation of transgenic animals.【Method】Family line was established with self-developed hLF transgenic goats (G0♂), F1 generation was composed by integrated offspring individuals of G0♂ mated with ordinary dairy goat , F2 was generated by F1 males mated with F1, F2 or ordinary females, and F3 was generated by F2 males mated with F2 females. PCR and ELISA were implied to identify the integration and expression of exogenous gene in offsprings. Reproductive data such as quantity of offspring with different mating patterns, rate of transgenic integrated individuals, conception rates, abnormal rates of lamb in each generation were counted statistically. 【Result】 After several reproductive cycles, no abnormalities of conception rates and lambing rates were observed. The results showed that, there were 19 hLF transgenic individuals in F1 generation, with integration rate of 24.7% (19/77), 86 individuals in F2 generation, in which 62 were generated by F1♂×N♀, 16 were obtained by F1♂×F1♀, and 8 were obtained by F1♂×F2♀, with the rate of 33.2% (62/187), 76.2% (16/21) and 34.8% (8/23), respectively, 9 hLF transgenic individuals in F3 generation and the integration rate of F3 was 52.9% (9/17). ELISA was conducted to measure the expression of hLF in F1, F2 and F3 in 20 weeks, and the results showed that hLF expressed stably in each transgenic generation and the average expression level in milk were 351.75, 340.00 and 326.25 μg•mL-1, respectively.【Conclusion】The results suggested that, there is no significant effect of transgenosis on reproductive performance of hLF transgenic goat, and exogenous hLF gene could inherit and stably express in each generation of goats.

Cloning and Expression of Cellular Retinoic Acid Binding ProteinⅠGene in Inner Mongolian Cashmere Goats

LI Hua, SU Li-Ning, LIU Dong-Jun, LI Xue-Feng, XU Ri-Gan

Scientia Agricultura Sinica. 2012, 45(13): 2743-2750. doi:10.3864/j.issn.0578-1752.2012.13.019

Abstract ( 758 )

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【Objective】The cDNA sequence of cellular retinoic acid binding protein I (CRABPⅠ) gene was cloned in Inner Mongolian cashmere goats, and the protein structure gene and expression were also analyzed. All these would establish a foundation for molecular mechanism of follicle and cashmere formation. 【Method】 The cDNA sequence of CRABP I gene was cloned by RT-PCR in Inner Mongolian cashmere goats. The protein structure was predicted through bioinformatics approach, the mRNA expression of the gene at four embryo ages in skin of cashmere goat were detected through real time PCR. 【Result】The length cDNA is 679 bp (JN936490) , its open reading frame (ORF) is 414 bp，which shares high similarity with other species. CRABPⅠprotein has no obvious signal peptide, transmembrane segments, N-glycosylation sites and O-glycosylation sites. The secondary structure of CRABPⅠprotein consisted of mainly β sheets, α helixes and loops, also few turn and coil. CRABPⅠ gene in cashmere goats was highly expressed on 90 d compared with on 100 d, 120 d and 130 d (P＜0.05). 【Conclusion】The open reading frame (ORF) of CRABPⅠgene is conserved among different species, but it has characteristics at 33 and 123 amino sites in cashmere goats. The level of mRNA expression was the highest on 90 d at four embryo stages, the polymorphism of this gene in different breeds and its regulation mechanism to sebaceous gland of hair follicle needs to be studied further.

Rapid Detection of Campylobacter jejuni Using Loop-Mediated Isothermal Amplification Assay

XU Yi-Gang, LI Dan-Dan, TIAN Chang-Yong, LIU Zhong-Mei, LI Su-Long, ZHANG Guo-Cai

Scientia Agricultura Sinica. 2012, 45(13): 2751-2757. doi:10.3864/j.issn.0578-1752.2012.13.020

Abstract ( 759 )

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【Objective】Campylobacter jejuni is a major zoonotic pathogen, mainly initiates its transmission with contaminated water, milk products and raw poultry. Thus, the development of a rapid and accurate diagnostic method is an important measure to prevent and control C. jejuni.【Method】Loop-mediated isothermal amplification (LAMP) assay was introduced to develop a rapid method for the detection of C. jejuni using the ORF-C sequence of C. jejuni as the target gene.【Result】Twenty-one bacterial strains were used to evaluate the specificity of the LAMP method. The result showed that only C. jejuni was positive and the other strains were negative in LAMP, indicating that the LAMP method was highly specific to the target pathogen with the sensitivity of 5CFU per test tube for C. jejuni cultivated and 9 CFU per test tube for its contaminated materials.【Conclusion】The LAMP method developed in this work is a sensitive, rapid and simple tool for the detection of C. jejuni.

Effects of Fertility-Promoting Powder on Trace Elements and Interleukin in Serum in Infertile Dairy Cattle

DENG Min, GU Xin-Li, LIANG Guang-Hua, LIU Xian-Xia, GAO Shu, XU Lei

Scientia Agricultura Sinica. 2012, 45(13): 2758-2763. doi:10.3864/j.issn.0578-1752.2012.13.021

Abstract ( 740 )

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【Objective】The law of trace elements (Cu, Fe, Mn, Zn) and interleukin (IL-1, IL-6) contents in serum using fertility-promoting powder to treat infertility caused by ovarian disease of dairy cattle were studied. 【Method】 Flame atomic absorption spectrometry and enzyme-linked immunization absorb assay were used to detect the levels of trace elements and interleukin contents in serum from the experimental and normal groups before and after treatment. 【Result】 After treatment, the effective rate of persistent corpus luteum was 91.67%, the effective rate of ovarian static was 88.89%. In persistent corpus luteum body, Cu, Zn contents were significantly higher than normal group in pre-treatment (P＜0.05) , Mn content was significantly lower than the normal group (P＜0.01), after treatment, their content levels closed to the normal group, and the difference was not significant (P＞0.05). The Fe, IL-1, and IL-6 contents closed to the normal group before and after treatment. In ovarian static body, Mn content was significantly lower than the normal group in pre-treatment (P＜0.01), IL-1 content was higher than the normal group (P＜0.05), after treatment, the contents were not significantly different (P＞0.05), but Cu,Fe,Zn, and IL-6 contents were not significantly different compared with the normal group before and after treatment (P＞0.05). 【Conclusion】The effect of fertility-promoting powder on trace elements and interleukin in serum can not be ignored in treatment of infertility caused by ovarian disease of dairy cattle.

Effects of Ryegrass Incorporation on Fertilizer N Cycling in a Double Rice System

ZHU Bo, YI Li-Xia, HU Yue-Gao, ZENG Zhao-Hai, TANG Hai-Ming, XIAO Xiao-Ping, YANG Guang-Li

Scientia Agricultura Sinica. 2012, 45(13): 2764-2770. doi:10.3864/j.issn.0578-1752.2012.13.022

Abstract ( 759 )

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【Objective】 Effects of N use efficiency and N cycling were examined when the fresh ryegrass residue was applied with urea in a double rice cropping system.【Method】 A 15N-labelled pot trail was conducted to estimate the effects of different treatments of urea (CF), half N from urea plus half N from ryegrass residue (RGCF) and the control (CK, no fertilizer) on soil mineral N concentration, rice yield, N use efficiency, N cycling and N2O emission.【Result】 Compared to CK, CF and RGCF increased the biomass dry matter and grain yield of double rice. In comparison with CF, RGCF increased rice biomass dry matter and grain yield by 5.9% and 7.3%, respectively. Compared to CK, CF and RGCF increased the total N accumulation of double rice. No significant difference in total N accumulation was found between RGCF and CF. In comparison with CF, RGCF increased N dry matter production efficiency, N grain production and N agronomic efficiency, reduced 15N recovery efficiency and 15N harvest index. When compared with CF, RGCF improved the proportion of 15N fertilizer residue in soil and decreased N2O emission.【Conclusion】Urea application in conjunction with ryegrass residue could increase N use efficiency and improve N cycling in double rice soils.

The Mechanism of Red Coloring Difference Between Skin and Cortex in Malus sieversii f. neidzwetzkyana (Dieck) Langenf.

WANG Yan-Ling, ZHANG Yan-Min, FENG Shou-Qian, SONG Yang, XU Yu-Ting, ZHANG You-Peng, CHEN Xue-Sen

Scientia Agricultura Sinica. 2012, 45(13): 2771-2778. doi:10.3864/j.issn.0578-1752.2012.13.023

Abstract ( 808 )

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【Objective】The aim of this study is to explore the red development mechanism by monitoring anthocyanin composition and content, transcriptional profile of anthocyanin related genes in Malus sieversii f. neidzwetzkyana (Dieck) Langenf.【Method】M. sieversii f. neidzwetzkyana (Dieck) Langenf was used as material to study anthocyanin composition of skin and cortex by HPLC, and to study the transcriptional profile of anthocyanin related genes by qPCR at four developmental stages. Anthocyanin contents in skin and cortex were determined at the four sampling times accordingly. 【Result】Cyanidin-3-galactoside was the main anthocyanin composition of skin and cortex, but anthocyanin content had different change trends at different developmental stages. The anthocyanin content of skin showed a declined pattern, while the anthocyanin content of cortex displayed an opposite tendency. Expression of the anthocyanin biosynthesis genes was correlated with anthocyanin content: expression of MsMYB10 in skin and cortex was very high, while expression of MsMYB10 showed an increasing pattern in skin and a ‘ high-low-high-low’ change trend in cortex during fruit development. 【Conclusion】The red coloring difference between skin and cortex of ‘Xiahongrou’ mainly relate to the anthocyanin content and the different expressions of anthocyanin gene. These results have provided a foundation for further exploring the red coloring mechanism and breeding cultivars of red-flesh apple.

Identification of the Secondary Follicle Cycling of Hexi Cashmere Goat

HE Yan-Yu, LUO Yu-Zhu, CHENG Li-Xiang, WANG Ji-Qing, LIU Xiu

Scientia Agricultura Sinica. 2012, 45(13): 2779-2786. doi:10.3864/j.issn.0578-1752.2012.13.024

Abstract ( 825 )

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【Objective】An experiment was conducted to identify the periodic change of ultrastructures of secondary follicle characteristics in the whole year, reveal the molecule regulation of growth of cashmere goat. 【Method】 A total of six cashmere goats of one year old were used in this experiment. The ultrathin slices of skin were made each month in the whole year, observed and photographed under transmission electron microscope after staining. 【Result】Following the development of down fiber, the ultrastructures of secondary follicle of Hexi cashmere goat showed a periodic change within a year. There were five different periods during the down fiber cycle. It was observed that the stages of telogen, pregrowing, anagen, precatagen and catagen were in January and February, March and April, May to August, September and October and November and December, respectively. 【Conclusion】 This study illustrated the five different stages of secondary follicle of Hexi cashmere goat within a whole growing cycle, and has provided detailed information on the research field of cashmere goat.

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