Scientia Agricultura Sinica

DNA-Marker Based Analysis of the Allelic Constitution and Subspecies Attribution of Super Hybrid Rice Yiyou 673

HUANG Ting-Xu, ZHU Yu-Jun, FAN Ye-Yang, YOU Qing-Ru, ZHOU Peng, YANG Dong, XIE Hua-An, ZHUANG Jie-Yun

Scientia Agricultura Sinica. 2013, 46(10): 1965-1973. doi:10.3864/j.issn.0578-1752.2013.10.001

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【Objective】 This study was conducted to determine the allelic constitution and subspecies attribution of the super hybrid rice Yiyou 673, so as to facilitate the utilization of beneficial alleles carried by Yiyou 673, and to provide information for establishing an appropriate indica-japonica proportion in rice breeding. 【Method】 Three major restorer lines of the three-line hybrid rice, including the founder parent Minghui 63, its derivative Minghui 86, and Fuhui 673 which is derived from Minghui 86, as well as cytoplasmic male sterile line Yixiang A, and Yiyou 673 which is developed by crossing Fuhui 673 to Yixiang A, were assayed with 17 markers for 13 genes of agronomically importance and 15 SSR markers for indica-japonica differentiation. 【Result】 Polymorphism was detected in eight of the gene marker regions, including two genes for heading date and yield traits, two key genes for grain quality, and four genes for blast resistance. At Wx locus which is responsible for amylose content and gel consistency of the milled rice, a high-quality allele was transmitted from Minghui 63 to Fuhui 673 through Minghui 86. At ALK locus controlling gelatinization temperature and two loci for blast resistance, Pib and Pita, the male and female parents of the F1 hybrid rice Yiyou 673 showed allelic complementation for high grain quality and resistance to blast. In addition, it was shown that Minghui 63, Minghui 86 and Fuhui 673 are all indica-inclined, but the japonica component of 40.0% in Minghui 86 and Fuhui 673 is higher than the value of 33.3% in Minghui 63.【Conclusion】 Beneficial alleles derived from Minghui 63 and other parental lines were pyramided in Fuhui 673, which were combined with those carried by Yixiang A to establish a favorable allelic complementation in Yiyou 673.

Establishment of Molecular Identity for Kenaf Germplasm Using SRAP Marker

LIU Qian, DAI Zhi-Gang, CHEN Ji-Quan, WEN Lan, GONG You-Cai, SU Jian-Guang

Scientia Agricultura Sinica. 2013, 46(10): 1974-1983. doi:10.3864/j.issn.0578-1752.2013.10.002

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【Objective】One hundred and twenty-seven accessions of kenaf gemplasm collected from different countries and areas, which contain cultivated, wild and related species, were selected and SRAP markers were used to establish molecular identity of kenaf germplasm.【Method】 Sequence- related amplified polymorphism(SRAP) marker was used for analyzing genetic relationships of the 127 different type accessions. Nei's similarity coefficients were generated and dendrogram was constructed using UMPGA by NTSYS-PC program.【Result】Three hundred and eighty-three bands were amplified by 40 SRAP primers, of which 375 were polymorphic and PPB was 97.9%. The cluster analysis based on UPGMA showed that the 127 accessions of kenaf germplasm were divided into four major groups at similarity coefficient of 0.70. All of the accessions could be effectively distinguished by specific bands and different primer combinations. The specific molecular identity for 127 accessions of kenaf germplasm were established with 99.99% probability of confidence.【Conclusion】A set of unique molecular identity was established for 127 kenaf germplasm based on 36 bands which were amplified by 15 core primer combinations.

Principal Component Analysis and Comprehensive Evaluation on Physiological Traits of Tolerance to Low Phosphorus Stress in Wheat Substitution

ZHENG Jin-Feng, MI Shao-Yan, JING Jiao-Jiao, BAI Zhi-Ying, LI Cun-Dong

Scientia Agricultura Sinica. 2013, 46(10): 1984-1993. doi:10.3864/j.issn.0578-1752.2013.10.003

Abstract ( 566 )

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【Objective】 The physiological indices of substitution lines in different phosphorus treatments were studied to select species of tolerance to low phosphorus stress. 【Method】 The comprehensive tolerance to low phosphorus stress of low phosphorus tolerance coefficient (LPTC) of soluble sugar, soluble protein content, SOD activity, POD activity, MDA content, leaf chlorophyll content, corticoid content, Pn, Tr, Gs, Ci, Fv/Fm of Chinese Spring (CS) - Synthetic 6x substitution lines as well as their parents were evaluated by principal components analysis and cluster analysis. 【Result】 The results showed that the 14 single indexes could be classified into 7 independent comprehensive components. The cluster analysis was used to divide the 21 substitution lines and their parents into three types of tolerance to low phosphorus. 2A substitution line and Synthetic 6x belonged to high tolerance types to low phosphorus stress; Chinese Spring and 4A, 7A, 2B, 1D and 3D substitution lines belonged to poor tolerance types to low- phosphorus stress. The others including 1A, 3A, 5A, 6A, 1B, 3B, 4B, 5B, 6B, 7B, 2D, 4D, 5D, 6D, and 7D substitution lines belonged to middle tolerance types to low- phosphorus stress.【Conclusion】 The comprehensive evaluation and principal component analysis provide a useful way for selecting species of tolerance to low phosphorus stress in wheat.

Effects of Intercropping with Waxy Maize on Crop Yield and Soil Properties of the Continuous Vegetable Field

ZHANG Jie-Ying, NING Tang-Yuan, FENG Yu-Peng, ZHANG Xue-Peng, SUN Tao, LI Zeng-Jia

Scientia Agricultura Sinica. 2013, 46(10): 1994-2003. doi:10.3864/j.issn.0578-1752.2013.10.004

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【Objective】The effects of soil properties and vegetable yield under different intercropping pattern treatments of waxy maize and maize straw returning were studied. Continuous cropping was used as a contrast. The aim was to supply academic base for solving continuous cropping obstacle and summer fallow problems in vegetable field. 【Method】 Broccoli, green sword bean and waxy maize were used as materials, and three treatments were set up. There were relay-intercropping mode of green sword bean and waxy maize, relay-intercropping mode of green sword bean and waxy maize with waxy maize straw returning, broccoli, green sword bean continuous cropping. The soil samples were used for chemical analysis after natural drying. Soil organic matter content, soil NH4+-N content, soil NO3--N content, the activity of urease, the activity of soil polyphenol oxidase and the activity of soil catalase were analyzed. At harvest period, the yields of broccoli, green sword bean and waxy maize were determined. 【Result】In the year 2010, the autumn vegetable yield of the relay-intercropping treatment was the highest, the autumn vegetable yield of the continuous cropping was the lowest. The principle was in agreement with the autumn vegetable yield of 2011. The results showed that soil organic matter content and soil urease activity were significantly increased under relay intercropping treatments and waxy maize straw returning treatment by using nutrients reasonably, compared with the vegetable continuous cropping. Intercropping or relay cropping decreased the inorganic nitrogen concentration, and increased the inorganic N utilizing rate in vegetable fields compared with sole cropping. Relay intercropping and waxy maize straw returning mode promoted microorganism propagation and made the soil urease activity at the high level. Vegetable continuous cropping caused soil nutrient over consumption and decreased the activity of urease. As more toxic substances from the root was released to the soil, this phenomenon made that the activity of soil polyphenol oxidase and catalase in vegetable continuous cropping fields were higher than that in other treatments, the increasing of the CAT activity will reduce the toxicity of the soil.【Conclusion】The results of the experiment indicated that crop yield increased effectively under relay-intercropping and straw returning. At the same time, relay-intercropping improved soil environment, organic matter content and urease activity increased, and the activity of soil polyphenol oxidase and catalase remained at an appropriate level.

Construction of a Highly Efficient Technology System in Anther Culture of Mutlifoliate Alfalfa

WU Zi-Nian, WEI Zhen-Wu, ZHENG Xi

Scientia Agricultura Sinica. 2013, 46(10): 2004-2013. doi:10.3864/j.issn.0578-1752.2013.10.005

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【Objective】 This experiment was carried out to establish an efficient anther culture system of mutlifoliate alfalfa, and get regenerate plantlet. It will also provide basic materials for the research of ploidy, genomics and molecular biology of alfalfa.【Method】 Firstly, Huaiyin alfalfa and a mutlifoliate alfalfa varieties named Grandeur were used as the parents to create a hybrid, and the anther of hybrid F1 was used as the explants. The ability of induction of callus after bud was studied by using the culture medium of NB added with 2,4-D (0.5 mg•L-1), NAA (0.3 mg•L-1) and KT (3.0 mg•L-1). The appropriate medium was screened for the differentiation of anther callus of alfalfa by orthogonal design L16 (45). In addition, the rooting ability from buds was studied by adding different concentrations of NAA into 1/2 B5 and 1/2 MS.【Result】The highest induction rate of alfalfa callus was in 15 to 28 days after budding, and the maximum value was 82.79%. The culture medium that was good for the differentiation of the callus of alfalfa anther was made up of BⅠwith CH (200 mg•L-1), 2-IP (0.5 mg•L-1), NAA (0.2 mg•L-1) and KT (1.0 mg•L-1), and the maximum regeneration rate was 54.05%. The best culture medium for adventitious bud was 1/2 B5 with NAA (0.3 mg•L-1), and the maximum rooting rate was 88.46%.【Conclusion】An efficient anther culture system of mutlifoliate alfalfa was established successfully, and the regenerated plants were obtained.

Identification of the Lipopeptides from Bacillus amyloliquefaciens Lx-11 and Biocontrol Efficacy of Surfactin Against Bacterial Leaf Streak

ZHANG Rong-Sheng, WANG Xiao-Yu, LUO Chu-Ping, LIU Yong-Feng, LIU You-Zhou, CHEN Zhi-Yi

Scientia Agricultura Sinica. 2013, 46(10): 2014-2021. doi:10.3864/j.issn.0578-1752.2013.10.006

Abstract ( 617 )

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【Objective】The objective of this study is to isolate and identify the lipopeptides produced by Bacillus amyloliquefaciens Lx-11 and their biocontrol efficacy against bacterial leaf streak (BLS). 【Method】 Three primers designed based on the known lipopeptide genes were used to amplify the corresponding genes from the genome of Lx-11 strain. Lipopeptides were identified by MALDI-TOF-MS techniques.【Result】The PCR products with three primer pairs were cloned and sequenced. The BLAST analysis showed that the sfp, fenB, ituA or bamA genes exist in the genome of B. amyloliquefaciens Lx-11. Three kinds of lipopeptides surfactin, bacillomycin D and fengycin were identified in the antimicrobial extract by the mass spectrum analysis. The antibacterial activity could be associated with surfactin-lipopeptides, and practically abolished in surfactin-deficient mutants. Lx-11 strain significantly reduced disease incidence of BLS.【Conclusion】The antibacterial activity of the surfactin- deficient mutant strain was practically abolished, which suggested that surfactin might play an important role in control of BLS.

Isolation and Mapping of NBS-LRR Resistance Gene Homology Sequences from Wheat

SHI Jing-Dong, ZHANG Xiao-Juan, HUANG Li-Li, HAN De-Jun, KANG Zhen-Sheng

Scientia Agricultura Sinica. 2013, 46(10): 2022-2031. doi:10.3864/j.issn.0578-1752.2013.10.007

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【Objective】The objective of this study is to isolate NBS-LRR class resistance gene homology fragments from wheat using homology-based method, and to validate the relationships with wheat stripe rust resistance gene Yr26.【Method】Primers were designed based on the conserved domains of the cloned plant disease resistance genes, and the nested PCR was used. The fragments were localized by Chinese Spring (CS) deletion lines of chromosome 1B, F2:3 populations derived from susceptible cultivar Avocet S and the lines 92R137 carrying Yr26 were used to evaluate the relationship between the isolated RGAs and Yr26. 【Result】Four open-reading resistance gene analogues (RGAs) were obtained, i.e., R105, R181, R326 and R405. The nucleotide sequences of the four RGAs were 369, 589, 528 and 618 bp. Homology research showed that the four fragments had typical conserved regions NBS-LRR, and the nucleotide identity of four fragments was from 24.35% to 38.36%. Only R405 was mapped in the deletion bin C-1BL-0.6-0.32 with Chinese Spring deletion lines and was co-segregated with a F2 population of 196 plants. It was initially hypothesized that the R405 might be the candidate sequence of Yr26.【Conclusion】In this study, four resistance homology fragments were obtained in the near isogenic lines (NILs) Nan137. R405 was mapped in the Yr26 region and co-segregated with Yr26, which provides a good prerequisite for further study.

Identification of Isolates of Verticillium Species from Cotton

ZHU He-Qin, FENG Zi-Li, LI Zhi-Fang, SHI Yong-Qiang, ZHAO Li-Hong, LI Cai-Hong, LIU Yi-Jie, WANG Ling-Fei

Scientia Agricultura Sinica. 2013, 46(10): 2032-2040. doi:10.3864/j.issn.0578-1752.2013.10.008

Abstract ( 525 )

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【Objective】In the genus of Verticillium, there are five ‘species’ as plant pathogens which could lead to plant wilt symptom. The aim of this study was to identify the specific ‘species’ caused Verticillium wilt of cotton.【Method】A total of 310 Verticillium isolates were obtained from 84 counties, 12 provinces of China. To clarify the ‘species’ of them, both observation on biological characteristics and molecular identification were used.【Result】A total of 298 isolates were typical V. dahliae producing black microsclerotia and verticillate conidiophores identified by specific PCR amplifications. However, 12 special isolates producing non-microsclerotia, black mycelia or chlamydospore were tested by temperature sensitivity, specific PCR amplification and ITS sequencing. Except for 2 V. nigrescens isolates, the rest 10 isolates were identified as V. dahliae, no V. albo-atrum was found. 【Conclusion】 In 3 main cotton growing regions of China, the causal agent of cotton Verticillium wilt is V. dahliae. And V. nigrescens also presented in Yangzi River and Yellow River basins. In cotton Verticillium wilt happening process, the interaction between V. dahliae and V. nigrescens needs to be further studied.

Effects of Nicotine on Feeding Behavior of Bemisia tabaci B and Q Biotypes Based on EPG and Liquid Diet Sac Technique

LI Xiao-Min, LI Jing-Jing, TANG Qing-Bo, YAN Feng-Ming

Scientia Agricultura Sinica. 2013, 46(10): 2041-2049. doi:10.3864/j.issn.0578-1752.2013.10.009

Abstract ( 529 )

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【Objective】 The effects of nicotine on the feeding behaviors of the Bemisia tabaci B and Q biotypes were investigated, with the aim of establishing a corresponding evaluating system based on the electrical penetration graph (EPG) and the liquid diet sac (LDS) technique. 【Method】 EPG method combined with a LDS was used to quantify the effect of the concentrations of nicotine on the feeding behaviors of the B. tabaci. 【Result】 Relative amplitudes and frequencies in EPG waveforms of the B. tabaci in 10% sucrose solution, compared to those in 5% and 15% sucrose solutions, were analogous to the waveforms of B. tabaci on plants. The durations of ingestion waveforms decreased with the increase of nicotine levels in 10% sucrose solution. Salivation (E1) and passive ingestion (E2) durations were reduced and the penetration duration was prolonged even at the low concentration (0.001%) of nicotine. The passive ingestion from LDS was completely inhibited at a higher concentration of nicotine (0.01%). Although the major EPG pathway variables between B. tabaci B and Q biotypes were similar, the duration of passive ingestion of Q biotype was significantly longer than that of the B biotype on the diet without nicotine or with lower nicotine concentration. 【Conclusion】 The valuable EPG variables for EPG+LDS system include the durations of C, E1 and E2 patterns. Ten percent sucrose solution in LDS was more suitable for recording the feeding behaviors of B. tabaci using EPG technique. The feeding behaviors of B. tabaci B and Q biotypes were signi?cantly inhibited by nicotine, but Q biotype had higher nicotine adaptability than B biotype.

Influence of Carbonized Maize Stalks on Fractions and Availability of Phosphorus in Brown Soil

GUAN Lian-Zhu, CHAN Zhong-Xiang, ZHANG Jin-Hai, ZHANG Guang-Cai, ZHANG Yun

Scientia Agricultura Sinica. 2013, 46(10): 2050-2057. doi:10.3864/j.issn.0578-1752.2013.10.010

Abstract ( 623 )

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【Objective】 The effects of carbonized maize stalks on the brown soil phosphorus fractions and availability were studied to ascertain whether or not the carbonized maize stalks is good for soil phosphorus fertility development.【Method】 Laboratory culture technique was used to compare the difference among the carbonized, non-carbonized and decomposed maize stalks treatments (same quantity carbon application in different maize stalks treatments), and the chemical fertilizers treatment alone with same amount application of chemical fertilizer. The fractions of soil phosphorus were determined by the modified Hedley’s sequential extraction procedure. The adsorption and desorption isotherms for soil phosphorus were determined by the method of Yan Chang-Sheng with Constant temperature shaking. 【Result】 The total phosphorus content in the soil could be significantly improved by all kinds of maize stalks treatments compared with the no maize stalks (single chemical fertilizer) treatment, and the phosphorus contents increased were significantly correlated with the quantities of maize stalks added into the soil. The soil phosphorus fractions could be affected significantly by the added different maize stalks. Although the available phosphorus content was not high in the carbonized maize stalks, but the sodium bicarbonate-extractable inorganic and organic phosphorus contents could be significantly improved by application of the carbonized maize stalks. The adsorption amount of phosphate ions could be reduced significantly by the maize stalks treatments, and the impact of carbonized maize stalks treatment was the most obvious. Whether the maize stalks was decomposed or not, the change of the phosphorus adsorption quantities was not obvious. 【Conclusion】 The application of carbonized maize stalks can promote the soil phosphorus availability, reduce soil phosphorus fixations, and improve the utilization ratio of the phosphorus fertilizer, but the increasing activity of soil phosphorus may also increase the phosphorus environmental risks.

Establishiment of Fertilization Recommendation Indexes of Rapeseed Soil Based on the “3414” Field Experiments in the Middle of Sichuan Hilly Regions

HUANG Yi, LI Ting-Xuan, ZHANG Xi-Zhou, JI Lin

Scientia Agricultura Sinica. 2013, 46(10): 2058-2066. doi:10.3864/j.issn.0578-1752.2013.10.011

Abstract ( 568 )

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【Objective】To provide a theoretical basis for balanced fertilization of rapeseed production in the middle of Sichuan hilly regions, abundance and deficiency indexes of soil nutrient and fertilization recommendation indexes were established. 【Method】Data were analyzed based on the “3414” experiments of rapeseed in the field from 2005 to 2009. Making use of plentiful-lack of soil nutrient as an index, soil available phosphorus (AP) and available potassium (AK) were divided into five levels on the basis of different relative yields. Nitrogen fertilizer application rate (N) on average was estimated based on the method of total quantity control, while phosphorus fertilizer (P2O5) and potassium fertilizer application rates (K2O) were calculated by the method of fertilizer response function. 【Result】 Regional optimal application rate of chemical fertilizer N on average is (170±50) kg•hm-2. Phosphorus fertilizer application rates (P2O5) are 100-120,70-100, 50-70, 30-50, ＜30 kg•hm-2 (AP＜5, 5-15, 15-25, 25-45 and ＞45 mg•kg-1), while potassium fertilizer application rates (K2O) are 90-110, 70-90, 50-70, 30-50, ＜30 kg•hm-2 (AK＜30, 30-60, 60-120, 120-240 and ＞240 mg•kg-1). 【Conclusion】Compared with the fertilization recommendation rate calculated by the original system, the fertilization recommendation rate above suggests that nitrogen and phosphorus fertilizer application rates should be decreased, while potassium fertilizer application rates ought to be increased.

Status of Soil Fertility in Vineyard-Producing Areas of Hebei Province

YIN Xing, JI Yan-Zhi, NI Yu-Xue, WANG Tan-Kui, ZHANG Li-Juan

Scientia Agricultura Sinica. 2013, 46(10): 2067-2075. doi:10.3864/j.issn.0578-1752.2013.10.012

Abstract ( 571 )

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【Objective】 Soil nutrient fertility status was investigated to formulate a suitable fertilization strategy so as to help to raise its economic and environmental benefits. 【Method】 Soil samples from different layers (0-20 cm, 20-40 cm and 40−60 cm) were collected from 99 vineyard producing areas located in Changli, Huailai, Xuanhua and Zhuozhou counties in Hebei Province, respectively. 【Result】The content of soil organic matter was low at large, the deficiency rate of 0−20 cm, 20−40 cm and 40−60 cm was 52%, 76% and 90%, respectively. Moreover, the CV of vineyards soil N, P and K was high and the CV of surface soil was 113.1%, 143.5% and 68%, respectively. Similarly, the concentrations of Fe and Cu in vineyards soil were too high, 94% and 99% in surface soil above suitable content, respectively. However, the contents of Zn and Mn were low and the deficiency rate of Zn and Mn was 56% and 100% in surface soil, respectively. 【Conclusion】It was suggested that more organic matter, Zn, Mn, rational application of nitrogen and less Cu fungicide should be used in the tested vineyards.

Effects of Exogenous Nitric Oxide and Abscisic Acid on Photosynthesis Characteristics in Pepper Seedlings Under Autotoxicity Stress

ZHANG Guo-Bin, YU Ji-Hua, FENG Zhi, MA Yan-Xia, 吕Jian

Scientia Agricultura Sinica. 2013, 46(10): 2076-2084. doi:10.3864/j.issn.0578-1752.2013.10.013

Abstract ( 626 )

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【Objective】 The aim of the experiment was to explore the effects of exogenous NO donor (SNP) and abscisic acid on pepper resistance ability of autotoxicity. 【Method】 Pepper cultivar ‘Longjiao 2’ was used as materials in this experiment to investigate the effects of SNP (150 μmol•L-1) and ABA(100 μmol•L-1) on gas exchange parameters, cholorophyll fluorescence parameters and chlorophyll content in pepper seedlings under autotoxicity stress. The seedlings of pepper were treated with soil and substrates extracts of 3 years continuous cropping pepper. 【Result】 It was found that all of the soil and substrates extracts reduced Pn, Gs, Tr, Fv/Fm, ΦPSⅡ, qP, chlorophyll content and ratio of chlorophyll a/b obviously, but increased Ci and NPQ significantly. Under autotoxicity stress, exogenous NO treatments significantly increased Pn, Gs, Fv/Fm, ΦPSⅡ, qP and chlorophyll content, but reduced Ci and NPQ significantly, and had no obvious influence on Tr. Exogenous ABA treatments significantly increased Pn, Fv/Fm, ΦPSⅡ, qP and chlorophyll content, but reduced Ci and NPQ significantly, and had no obvious influence on Gs and Tr under autotoxicity stress.【Conclusion】The reduction in net photosynthesis rate caused by autotoxicity stress was considered to be a result of non-stomatal restriction mainly, and autotoxicity on pepper treated with soil extracts was serious greatly than substrates extracts. Under autotoxicity stress, exogenous NO and ABA application increased chlorophyll content, maintained PSII center activities and photosynthetic capacity, and enhanced autoxicity tolerance in pepper.

Developments of Molecular Markers Tightly Linked to Fon-1 for Resistance to Fusarium oxysporum f. sp. niveum Race 1 in Watermelon

ZHANG Yi, ZHANG Hai-Ying, GUO Shao-Gui, REN Yi, ZHANG Jie, GENG Li-Hua, LIANG Zhi-Huai, XU Yong

Scientia Agricultura Sinica. 2013, 46(10): 2085-2093. doi:10.3864/j.issn.0578-1752.2013.10.014

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【Objective】 In order to provide technical support for molecular marker-assisted breeding for Fusarium wilt resistance in watermelon, molecular markers tightly linked to Fon-1 for Fusarium oxysporum f. sp. niveum race 1 resistance were developed.【Method】An F2 segregating population derived from a cross between the cultivated resistant male parent ‘Calhoun Gray’ and the susceptible female parent ‘Black Diamond’. The bulked segregant analysis (BSA) was employed to map the Fon-1 gene interval preliminaryly. A series of candidate single nucleotide polymorphism (SNP) sites that are linked to the target traits were obtained, and resequencing analysis within the target gene interval among 11 cultivated watermelon accessions was conducted. And then, the cleaved amplified polymorphic sequence (CAPS) or derived cleaved amplified polymorphic sequence (dCAPS) markers were designed to verify the linkage relationship between these CAPS/dCAPS marker and Fon-1 gene using F2 segregating population and germplasm resources. 【Result】 The preliminary results from screening of the resistant and susceptible DNA pools with 950 pairs of SSR/Indel/dCAPS primers mapped the Fon-1 gene to a 15 cM interval of watermelon chromosome 1. Three markers, 7716_fon, 7419_fon and 4451_fon, which were demonstrated to be tightly linked to Fon-1 gene, were developed using the SNP sites confirmed in the F2 population and 164 cultivars and the genetic distance between the markers and the Fon-1 gene was estimated to be 0.8 cM, 1.0 cM and 2.8 cM, respectively.【Conclusion】Three CAPS/dCAPS markers that can effectively distinguish the resistance/susceptibility to Fusarium oxysporum f. sp. niveum race 1 in cultivated watermelon were developed in this study and these markers will provide useful technical support for molecular breeding for resistance against F. oxysporum f. sp. niveum race 1 in cultivated watermelon.

Molecular Cloning and Preliminary Functional Analysis of Domains of Duck Retinoic Acid Inducible Gene I

CHEN Yang, HUANG Zheng-Yang, ZHANG Yang, LI Xin-Yu, ZHEN Ting, WU Ning-Zhao, XU Qi, CHEN Guo-Hong

Scientia Agricultura Sinica. 2013, 46(10): 2094-2102. doi:10.3864/j.issn.0578-1752.2013.10.015

Abstract ( 504 )

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【Objective】Duck RIG-I (duRIG-I) gene was cloned and the functions of its different domains were analyzed preliminarily. 【Method】 The CDS of duRIG-I gene was cloned on the basis of the sequence submitted to GenBank with RT-PCR and was analyzed by bioinformatics. The eukaryotic expression vectors of N-terminal, C-terminal and whole-length of duRIG-I gene with 6*his tags were constructed to transfect DF1, and then the transcription and expression of the three recombinant plasmids in cells were detected via RT-PCR and indirect immunofluorescent assay, respectively. Meanwhile, the expressions of chicken IFN-β, Mx1 and PKR mRNA were detected by real-time PCR.【Result】The whole-length of duRIG-I CDS was 2 802 bp encoding 933 amino acids. All the recombinant protein of duRIG-I could express normally in DF1. The results of RT-qPCR indicated that CARDs significantly up-regulated the mRNA level of IFN-β, Mx1 and PKR genes.【Conclusion】The various domain fragments of duRIG-I express normally in DF1. The N-terminal of duRIG-I plays a vital role in regulating the expression of downstream genes of RLR antiviral signal pathway.

Expression and Localization of Claudin-11 in Adult Alpaca Ovary

GENG Jian-Jun, GUO Qing-Yun, PANG Ya-Miao, BAI Jun-Ming, HE Jun-Ping

Scientia Agricultura Sinica. 2013, 46(10): 2103-2109. doi:10.3864/j.issn.0578-1752.2013.10.016

Abstract ( 488 )

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【Objective】To explore the mechanism of Claudin-11 in promoting the follicular development and regulating the ovulation, an experiment was carried out to investigate the expression and localization of Claudin-11 in adult alpaca ovary. 【Method】 Using the adult female alpaca as the object of study, the expression of Claudin-11 in the ovary was analyzed by RT-PCR and real-time PCR. Immunohistochemistry and western blotting were used to explore the localization of Claudin-11 in alpaca ovary.【Result】The expression of Claudin-11 in ovarian primary follicle was weaker than in the secondary follicle. Polyclonal antibody happened specific immune response with the crude extract from the ovarian. The expression of Claudin-11 in ovarian primary follicle and secondary follicle was weaker than that in flat or cube follicle cell cytoplasm. Claudin-11 was mainly located in the granulosa cell of the secondary follicle endothecium.【Conclusion】The results showed that Claudin-11 expressed in adult alpaca ovary, and there was a specific cellular localization.

Study on the Cloning of Coding Sequence and Ontogenetic mRNA Expression of Ovine UCP4 Gene

ZHOU Sha-Sha, LIU Bao-Feng, WEI Lin-Lin, WANG Jing-Lin, LIU Jian-Hua, LIANG Chen, QIAO Li-Ying, LIU Wen-Zhong

Scientia Agricultura Sinica. 2013, 46(10): 2110-2118. doi:10.3864/j.issn.0578-1752.2013.10.017

Abstract ( 438 )

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【Objective】This study aims to clone the coding sequence (CDS) of ovine UCP4 gene, to analyze the CDS and its coding protein structures, and to profile the ontogenetic mRNA expression so that to lay a theoretical foundation for future research on the structure and function of this gene. 【Method】The CDS was amplified from ovine cerebrum tissue by reverse transcription- polymerase chain reaction (RT-PCR). Physicochemical features and structural characteristics of ovine UCP4 were analyzed by bioinformatics methods. Ninety-six animals from two sheep breeds with significant differences in tail types (Guangling Large Tailed- and Small Tailed-Han Sheep) were used to study the ontogenetic mRNA expression by real-time quantitative PCR in eight tissues (cerebrum, cerebellum, hypothalamus, pituitary, and subcutaneous fat, perirenal fat, mesenteric fat and tail fat) from 2-to 12-month old age at two-month interval. 【Result】The sequencing data showed that the length of complete CDS was 972 bp encoding a protein of 323 amino acids. The isoelectric point (pI) of the protein was 9.43, and the molecular weight was 35.73 kDa. The percentages of helix, strand and loop were 56.04%, 7.12% and 36.84%, respectively, in the secondary structure. This transmembrane protein had no signal peptide but with two predicted glycosylation sites and 15 potential phosphorylation sites. The UCP4 mRNA expressed in all these tissues, but overexpressed in brain tissue other than in adipose tissue. Generally, breed, tissue and the months of age had significant influences on the UCP4 mRNA expression. 【Conclusion】 The obtained whole CDS of ovine UCP4 gene, expression profile and its limiting factors are of scientific significance to further studies on the relationship between the gene structure and energy metabolism.

Mapping QTL for Heat Tolerance at Grain Filling Stage in Common Wheat

LI Shi-Ping, CHANG Xiao-Ping, WANG Cheng-She, JING Rui-Lian

Scientia Agricultura Sinica. 2013, 46(10): 2119-2129. doi:10.3864/j.issn.0578-1752.2013.10.018

Abstract ( 656 )

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【Objective】In this study, a doubled haploid (DH) population with 150 lines, which was derived from the cross of two Chinese common wheat cultivars Hanxuan 10 and Lumai 14, was used as the plant material to identify the heat tolerance index (HTI) for related physiological traits and thousand-grain weight (TGW) at grain filling stage in common wheat and carry out quantitative trait loci (QTL) analysis. The purpose was to identify the essential QTL with stable and remarkable effects and find a theoretical basis on marker-assisted selection for improving the heat tolerance in wheat breeding program. 【Method】 The mixed-model-based composite interval mapping method was employed to identify QTL for HTI of related traits in two soil moisture environments. 【Result】A total of 12 additive QTL and 17 epistatic QTL for HTI of TGW and physiological traits related to heat tolerance were located on all chromosomes except 1D, 6D and 7B under two soil moisture conditions. One single additive QTL can explain 2.64%-11.41% phenotypic variance for HTI, and a pair of epistatic QTL can explain 2.45%-8.84% phenotypic variance for HTI. Nine of 12 additive QTL have interaction effects with the environments, and the interaction effect of single additive QTL with the environment can explain 1.41%-4.66% phenotypic variance for HTI. Only 5 of the 17 epistatic QTL exist interaction effects with the environments, and the interaction effect of single pair of QTL with the environment can explain 0.62%-2.32% phenotypic variance for HTI. The allelic contribution to the HTI QTL came from both parents. Some DH lines were more tolerant to the heat stress than their parents.【Conclusion】 To evaluate the heat tolerance at grain filling stage, the HTI for TGW is a direct criteria, the following two physiological index are indirect criteria, i.e. the HTI for flag leaves relative water content (IRWC) is an available index under rainfed condition, and the HTI for canopy temperature depression (ICTD) is suitable under well-watered condition. The QTL for HTIs mainly distribute on the chromosomes 1B, 2D, 5A, 5B, 6A, 6B and 7A, showing that these chromosomes have close relationship with heat-tolerance at grain filling stage. The genetic effects of QTL for TGW HTI and chlorophyll content (CC) mainly are additive effect, and that of CTD and chlorophyll fluorescence parameters (CFP) are mainly epistatic effect. The additive effect and the epistatic effect are almost equal in the genetic effect of the QTL for the RWC HTI.

Effects of Cultivation Methods on Yield and Utilization of Temperature and Light of Late Japonica Rice in Southern Double Cropping Rice Areas

ZHANG Jun, ZHANG Hong-Cheng, HUO Zhong-Yang, LI Guo-Ye, DONG Xiao-Bo, HUA Jin, GUO Bao-Wei, ZHOU Pei-Jian, CHENG Fei-Hu, HUANG Da-Shan, CHEN Zhong-Ping, CHEN Guo-Liang, DAI Qi-Gen, XU Ke, WEI Hai-Yan, GAO Hui

Scientia Agricultura Sinica. 2013, 46(10): 2130-2141. doi:10.3864/j.issn.0578-1752.2013.10.019

Abstract ( 506 )

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【Objective】 Under the conditions of late rice of southern double cropping systems, five different cultivar types of japonica were used to investigate the effects of cultivation methods on yield, growth period and utilization of temperature and light. 【Method】 In Shanggao of Jiangxi province, four types of experimental materials including late-maturing medium japonica rice, early-maturing late japonica rice, medium-maturing late japonica rice and late-maturing late japonica rice as well as three cultivation methods including hand transplanting (HT), cast transplanting(CT) and mechanical transplanting (MT) were used to comparatively study the effect of different cultivation methods on the yield, growth period and utilization of temperature and light. 【Result】 The yields of different types of late japonica rice all demonstrated that HT>CT>MT, and different significant or very significant levels were reached between the yields of different cultivation methods. Compared with the HT, the yields were reduced by 6.05%-25.70% and 9.94%-39.04% under CT and MT, respectively. These differences resulted from the total spikelet, seed setting rate and grain weight under HT were higher than under CT and MT. Compared with the HT, the growth periods of CT and MT were delayed, and their total growth periods were significantly shortened by 4.9 and 10.3 days. For different types of varieties, the shortened day of late japonica was more than that of medium japonica. For the same mature period types, the shortened day of japonica hybrid rice was more than that of japonica conventional rice. The utilization efficiency of accumulated temperature and light time of various types of varieties at different developmental stages showed that HT>CT>MT, and the whole growth period trended similarly. The utilization efficiency of temperature and light in the whole growth period under MT was only 85.66% and 80.75%. 【Conclusion】On the basis of the above results, a preliminary planning for the suitable range and its supporting suitable variety types of different cultivation methods was made. HT, CT and MT were all suitable in this area, but each cultivation method had its suitable variety types.

Effect of Irrigation and Nitrogen Application Regimes on Senescent Characters of Roots and Leaves in Cotton with Under-Mulch-Drip Irrigation

LUO Hong-Hai, ZHANG Hong-Zhi, TAO Xian-Ping, ZHANG Ya-Li, ZHANG Wang-Feng

Scientia Agricultura Sinica. 2013, 46(10): 2142-2150. doi:10.3864/j.issn.0578-1752.2013.10.020

Abstract ( 561 )

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【Objective】 To investigate the effect of irrigation and nitrogen application regimes on the senescent characters of roots and leaves, yield and water use efficiency (WUE), experiments were conducted using cotton with under-mulch-drip irrigation at yield forming stages. Providing evidence for regulating cotton senescence and improving yield will be of great significance to the cotton production.【Method】All the cotton plants were planted in soil columns under the same conditions but with different amounts of water and nitrogen supply at different growth stages, and different distribution proportions, before inseminating. There were eight treatments of four amounts of water and two amounts of nitrogen used to research the effect of water and nitrogen application regime on root growth, root vigor, the activity of protective enzyme in roots and leaves, yield and WUE of cotton.【Result】The optimum water and nitrogen application regime that irrigated before insemination and limited irrigation before full flowering stage, followed by adequate recovery irrigation along with fertilization again at flowering and boll stage was the best. This treatment promoted root growth in deeper soil layers, strengthened root vigor, increased superoxide dismutase (SOD) activity in roots and leaves, and decreased malondialdehyde (MDA) content so that the senescence processes were delayed. Furthermore, it increased distribution from photosynthetic production to production in reproductive organs and increased boll weight. Consequently, a comparatively high yield and WUE were gained in this treatment.【Conclusion】The most efficient water and nitrogen application regime is that decrease water and nitrogen supply before full flowering stage, but increase distribution proportion of water and nitrogen at the middle and late growth stages. This can delay early senescence of cotton, promote distribution from photosynthetic production to reproductive organ production, increase cotton yield with under-mulch-drip irrigation, and increase the WUE.

The Relationship of Grape Leaf Stomatal Conductance and Water Potential with Leaf Position Under Drought Conditions

WANG Jing-Jing, MO Wei-Ping, JIA Wen-Suo, LIU Guo-Jie

Scientia Agricultura Sinica. 2013, 46(10): 2151-2158. doi:10.3864/j.issn.0578-1752.2013.10.021

Abstract ( 585 )

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【Objective】 An experiment was conducted to study the regulations of the changes of stomatal conductance and water potential at different leaf positions under drought conditions.【Method】The stomatal conductance and water potential at different grape leaf positions were detected by 3005F01 Plant Water Status Console and SC-1 Leaf Porometer after drought treatments.【Result】Under well-watered condition, stomatal conductance had a positive linear correlation with the leaf position; However, a slight drought could reduce the stomatal conductance at upper leaf position significantly, with not much influence on lower leaf position. A contrast phenomenon was found under severe drought condition, the stomatal conductance declined with the increase in leaf position, while the water potential still had a positive correlation with leaf position.【Conclusion】No relationship between stomatal conductance and water potential under well-watered condition was found. The sensitivity to drought of leaves at different leaf positions had an obvious difference, which was closely related to the change of water potential and corresponding feedback regulation.

Evaluation of the Effect of Lentinus edodes Powder on Dough Rheological Properties with Mixolab

HU Qiu-Hui, GAO Yong-Xin, YANG Wen-Jian, FANG Yong, MA Ning, YUAN Biao, ZHAO Li-Yan

Scientia Agricultura Sinica. 2013, 46(10): 2159-2167. doi:10.3864/j.issn.0578-1752.2013.10.022

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【Objective】Rheological properties of dough with different amount of Lentinus edodes powder was investigated, which will lay a foundation for development of functional Lentinus biscuits as functional material.【Method】Mixolab was employed to investigate the effects of Lentinus edodes powder through 60 mesh sieve on rheological properties of dough, such as absorption, mixing, gluten index, viscosity, amylase activity, retrogradation, and gelatinization characteristics.【Result】Results showed that, in the range of 5%-20%, with the increase of Lentinus edodes powder, the dough viscosity and heat stability were significantly reduced, while starch gelatinization was increased, which resulted in the smaller stoma, product thickness and the volume.【Conclusion】 The effect of Lentinus edodes powder on the rheological properties was significant when the addition was higher than 15%, and ten percents of Lentinus edodes powder was known as the optimal addition amount for the product of biscuits. Meanwhile, it improved the nutritional value of biscuits greatly, and strengthen the nutritional content of Ca, Fe, Zn and lentinan.

Effect of Lycium Barbarum Polysaccharide on Production Performance and Serum Biochemical Indexes in Lactating Rabbit

CHEN Sai-Juan, LIU Ya-Juan, WANG Yuan-Yuan, CHEN Dan-Dan, LIU Tao, GU Zi-Lin

Scientia Agricultura Sinica. 2013, 46(10): 2168-2174. doi:10.3864/j.issn.0578-1752.2013.10.023

Abstract ( 673 )

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【Objective】This study was performed to investigate the effects of lycium barbarum polysaccharide (LBP) on production performance and serum biochemical indexes in lactating rabbit.【Method】A total of 48 lactating rabbits in the same period of parturition were selected and randomly divided into 4 groups for 1-monthtrial. GroupⅠ was used as a control group and the rabbits were fed with basal diet. Group Ⅱ, Ⅲ, and Ⅳ were used as experimental groups and the rabbits were fed with basal diet plus 0.5%, 0.75%, and 1% LBP.【Result】The results showed that compared to the control group, 35-day-old weaned rabbit weight gain of experimental groups Ⅱ, Ⅲ, and Ⅳ increased by 5.95%, 4.60% and 6.47%, respectively (P＞0.05). The weaning survival rate of experimental groups Ⅱ, Ⅲ, and Ⅳ was significantly improved (P＜0.01). The female rabbit lactating capacity and average daily feed take of experimental groups Ⅱ, Ⅲ, and Ⅳ were not different significantly(P＞0.05). The lactating rabbit weight loss of experimental groups Ⅱ, Ⅲ, and Ⅳ decreased by 20.9%, 16.2% and 15.5%, respectively (P＞0.05). The level of LDH in experimental groups Ⅱ, Ⅲ, and Ⅳ was, respectively, decreased by 32.6% (P＜0.01), 19.8% (P＜0.05) and 48.3% (P＜0.01). The level of the other serum biochemical indexes in experimental groups Ⅱ, Ⅲ, and Ⅳwas not different significantly(P＞0.05). 【Conclusion】It was concluded that adding appropriate LBP in lactating rabbit diet could improve newborn rabbits weaning survival rate, promote newborn rabbits growth, reduce lactating rabbit weight loss, and no adverse effects on serum biochemical indexes.

Cloning and Expressing Glideosome Associated Protein Genes of Babesia motasi and Preparation of Their Polyclonal Antibodies

WANG Jin-Ming, LIU Ai-Hong, REN Qiao-Yun, MA Mi-Ling, LIU Zhi-Jie, LI An-Yan, LI You-Quan, ZHAO Shuai-Yang, ZHANG Hao-Hao, YIN Hong, LUO Jian-Xun, GUAN Gui-Quan

Scientia Agricultura Sinica. 2013, 46(10): 2175-2182. doi:10.3864/j.issn.0578-1752.2013.10.024

Abstract ( 437 )

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【Objective】To prepare polyclonal antibodies of component genes from glideosome, gap45, gap50, myo A, and mlc were cloned and expressed in prokaryotic system. 【Method】 Rapid amplification of cDNA end (RACE) was used to amplify the full lengths of gap45, gap50, myo A, and mlc. The partial or complete open reading frames (ORF) of those genes were further inserted into prokaryotic expression vector and then transformed into BL21 (DE3) pLys to express by inducing with IPTG. The purified fusion proteins, with MagneGST™ Protein Purification System, were injected into rabbits to produce polyclonal antibodies. Specificities and titers of the polyclonal antibodies were determined by ELISA assay. 【Result】The full-length cDNAs of gap45, gap50, myo A, and mlc, were 664, 1 391, 2 538, 773 bp and open reading frames (ORF) were 567, 1 194, 2 514, and 606 bp in size. The polyclonal antibodies prepared had good specificities and the titers were more than 1:25 600.【Conclusion】In the present study, the full length of four component genes of B. motasi glideosome were amplified and their polyclonal antibodies were obtained, which will provide important biological materials for screening and identifying candidate antigens of vaccination and diagnosis, drug targets as well studying invasion mechanism of Babesia in the future.

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