Table of Content

12 October 2009, Volume 35 Issue 10

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Obtainment of Transformed Maize with Dehydration-Responsive Transcription Factor CBF4 Gene

Acta Agron Sin. 2009, 35(10):  1759-1763.  doi:10.3724/SP.J.1006.2009.01759

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The expression vector of CBF4 gene cloned from Arabidopsis thaliana by PCR and driven with promoterof rd29A gene, a preceding stress responsive gene, was constructed. Immatureembryo and embryogenic callus of several excellent maize inbred lines were transformed with biolistic bombardment. After in vitro culture steps of callus induction, resistance selection, differentiation and plant regeneration, thirty six transgenic plants were obtained. PCR, PCR-Southern and Southern blot analysis indicated that successful integration of alien target gene into genome of transgenic maize lines was happened. Measurement of drought physiological indexes under artificial drought treatment showed that either proline content or chlorophyll content of a transgenic line increased over one time than that of its wild type, suggesting that the drought resistance ability of transgenic maize might be improved to a certain extent.

Alternative Splicing of Photoperiod Response Gene Ppd-B1 in Wheat

GUO Zhi-Ai,ZHAO Guang-Yao,REN Zheng-Long,Jia Ji-Zeng

Acta Agron Sin. 2009, 35(10):  1764-1770.  doi:10.3724/SP.J.1006.2009.01764

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Alternative splicing creates multiple types of mRNA transcripts from a single gene, and can contribute to the regulation of gene function and protein diversity in eukaryotic cells. Photoperiod response plays a major role in controlling the heading time, yield, and adaptability in wheat (Triticum aestivum L.), and Ppd-B1 has been considered to have a potential effect on wheat responses to photoperiod changes. However, its alternative splicing has not been reported yet. In the present study, a photoperiod insensitive variety Chinese Spring carrying Ppd-B1 and another photoperiod sensitive variety Marquis carrying ppd-B1 were grown under four different photoperiod conditions. The cDNA and genomic DNA sequences of Ppd-B1 were isolated and characterized. The length of Ppd-B1 coding region was 3 053 bp with eight exons whose total size was 1 995 bp. The Ppd-B1 mRNA was alternatively spliced, producing multiple types of transcripts. There were three alternative splicing sites located in 5' UTR, exon 5, and intron 6, whose action led to exon increasing, alternative 5' end processing and intron retention. The alternative processing events that were augmented by the two former splicing sites remained the conserved Psedo-response regulator (PRR) domain in the deduced protein, whereas that directed by the third site resulted in frameshift mutation. Among the eight different types of Ppd-B1 transcripts produced by alternative splicing, four types (type a to d) expressed at relatively high levels with the frequency ranging from 32.6% (type d) to 13.0% (type b). They could be translated into full length proteins, and were likely to be functional. The remaining types (type e to h) expressed at very low levels with the frequency varying from 2.2% (type g) to 6.5% (type e). They gave rise to truncated peptides upon conceptual translation. The relative abundance of the different types of Ppd-B1 transcripts were changed by the photoperiod response characteristics of wheat varieties and the photoperiod conditions under which wheat plants were cultured. The data collected here paves the way for further studies to reveal the physiological function of Ppd-B1 alternative splicing in wheat.

Transgenic Ramie with Bt Gene  Mediated by Agrobacterium tumefacien and Evaluation of Its Pest-Resistance

FU Jia-Ping,WANG Bo,LIU Li-Jun,YANG Jin-Yu,WANG Xu-Xia,XING Xiu-Long,PENG Ding-X

Acta Agron Sin. 2009, 35(10):  1771-1777.  doi:10.3724/SP.J.1006.2009.01771

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Agrobacterium tumefaciens-mediated approach is an effectively and widely used to introduce foreign DNA into plants. However, there were few reports published on ramie transformation mediated by Agrobacterium tumefaciens up to date. In the present study, transgenic ramie containing Bt gene were obtained by our highly efficient Agrobacterium-mediated transformation system using ramie Luzhuqing. Three T₀ plants of transgenic ramie were chosen for further experiments and were transplanted into the fields. Laboratory assay of artificial infestation and the survey on agronomic traits and quality traits of T₀ plants were conducted. The results showed that T₀ transgenic plants exhibited more excellent pest resistance than the controls, and some were significantly superior to the controls. In addition, T₀ transgenic plants maintained the main agronomic traits, such as plant height, stem diameter, rate of effective tiller, raw fiber weight, fiber production ratio from stem and fiber production ratio from phloem, and quality traits, such as strength, filament breaking tenacity, fiber fineness, fineness uniformity and glue content of raw fiber in Luzhuqing. PCR and Southern blotting analysis showed that T₁ plants contained Bt gene, which indicated that Bt gene could be stably inherited. T₁ plants in the fields performed highly pest resistance compared with non-transgenic plants.

Phylogenetic Relationship of Ramie and Its Wild Relatives Based on Cytogenetics and DNA Sequences

LIAO Liang,LI Tong-Jian,LIU Zhong-Lai,DENG Hui-Sheng,XU Ling-Ling,PAN Qi-Hui3，

Acta Agron Sin. 2009, 35(10):  1778-1790.  doi:10.3724/SP.J.1006.2009.01778

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There are 31 species and 12 varieties of Boehmeria Jacq. in China. Boehmeria nivea is an important fiber crop in China and even in East Asia, which has been cultivated for about 4700 years. Because of the dispute in systematic relationship of different sections of Boehmeria among different scholars，the utilization or protection of B. nivea and its wild resources are restricted. In this paper, the karyomorphology of interphase nuclei and chromosome numbers were investigated in 19 species and 5 varieties, while the sequences of nuclearITS and chloroplast trnL-F were analyzed in 18 species and 9 varieties to explore the relationship between B. nivea and its wild relatives, to speculate how and where B. nivea originated and to effectively utilize plant resources of wild ramie. The cytological observation showed that the interphase karyomorphology of 24 taxa of Boehmeria should belong to pro-chromosome type and diffuse type according to the classification system of Tanaka, in which the interphase nuclei of Sect. Tilocnide and Sect. Boehmeria were pro-chromosome type and those of Sect. Zollingeriana, Sect. Phyllostachys and Sect. Duretia were pro-chromosome type and diffuse type. Cytogenetics examinatation of B. nivea and its 23 wild relatives (chromosome numbers of 15 taxa were reported in this paper at  the first time) showed that 19 taxa were diploid (2n=28), while three taxa in Sect. Duretia, were triploid (2n=42), one was tetraploid (2n=56) and one was pentaploid (2n=70). Analyses based on ITS and trnL-F sequences indicated that Boehmeria could be divided into three basic branches, A (including A₁ and A₂ sub-branch), B (including B₁ and B₂ sub-branch) and C. Different individuals or clones of B. clidemioides var. diffusa were gathered into different clades, indicating the hybridization and reticulate evolution among species in Boehmeria, andintrogressionbetween B. nivea and B. clidemioides var. diffusa. The wild core species resources of ramie should include both B. nivea and its three varieties and B. malabarica var. leioclada, B. clidemioides var. diffusa, which all should be intensively protected. It was inferred from the results two evolution routes of Boehmeria were: (1) pro-chromosome evolution line: A₂ originated from A₁, that is, Sect. Tilocnide from Sect. Boehmeria, and (2) diffuse evolution line: C originated from B, the order of origination is Sect. Boehmeria→Sect. Zollingeriana→Sect. Phyllostachys→Sect. Duretia. These two evolution routes may both originated from the same ancestor. It was implied based on the morphological traits of interphase nucleus and molecular experiment that the relationship of Sect. Zollongerie, Sect. Phyllostachys and Sect. Duretia was close, and the systematic relationship of these three sections based on morphological traits was not supported by the experimented evidences in cytology and molecular phylogeny in this study.

Molecular Mapping of Powdery Mildew Resistance Gene MlWE18 in Wheat Originated from Wild Emmer(Triticum turgidum var. dicoccoides)

HAN Jun,ZHANG Lian-Song,LI Gen-Qiao,ZHANG Hong-Tao,XIE Chao-Jie,YANG Zuo-Min

Acta Agron Sin. 2009, 35(10):  1791-1797.  doi:10.3724/SP.J.1006.2009.01791

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Wild emmer (Triticum turgidum var. dicoccoides) is an important germplasm for wheat improvement especially for resistance to powdery mildew caused by Blumeria graminis f. sp. tritici. Common wheat line 3D249 (Jingshuang 27//Yanda 1817/WE18/3/Wenmai 4, F₇), a derivative of wild emmer accession WE18 and susceptible elite common wheat lines, was found highly resistant to prevailing powdery mildew isolate E09 at both seedling and adult plant stages in Beijing, China. Genetic analyses of the F₁, F₂ segregating population and their F₃ progenies derived from a cross between susceptible line Xuezao and resistant line 3D249 indicated that the powdery mildew resistance of line 3D249 was controlled by a single dominant gene, temporarily designated MlWE18. By bulked segregant analysis (BSA), four SSR markers (Xwmc525, Xwmc273, Xcfa2040, and Xcfa2240), one RFLP-derived STS marker (Xmag1759) and one EST-STS marker (XE13-2) were found to be linked to MlWE18, with an order of Xwmc525–Xcfa2040–Xwmc273–XE13-2–Xmag1759–MlWE18–Xcfa2240 in the genetic linkage map. Using Chinese Spring nullisomic-tetrasomics, ditelosomics, and deletion lines, MlWE18 was physically mapped on chromosome 7AL terminal bin 7AL 16–0.85–1.00. However, the allelism of wild emmer derived MlWE18 to known powdery mildew resistance genes Pm1, PmU, MlIW72, Mlm2033, and Mlm80, all located on the same chromosome bin, need to be characterized in the future. The common wheat powdery mildew resistance line 3D249 provides useful new germplasm for disease resistance genes pyramiding and marker-assisted selection (MAS) in wheat breeding program.

Expression of Auxin-Related Genes during Dedifferentiation of Mature Embryo in Wheat

CHEN Jun-Ying,MA Ping-An,ZHAO Yi-Dan,ZHU Xue-Ping,CUI Yan,ZHANG Yan-Min

Acta Agron Sin. 2009, 35(10):  1798-1805.  doi:10.3724/SP.J.1006.2009.01798

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The quality of dedifferentiation determines the capacity of callus regeneration. Mature embryo is considered to be of the most potential for genetic transformation, but its transformation efficiency is still lower now.To elucidate the mechanism of mature wheat embryo dedifferentiation, we studied the expression profile of genes in dedifferentiation using Affymetrix microarray technique in mature embryos of wheat cultivar Yumai 18 cultured on MS medium supplemented with 2 mg L^-1 of 2,4-dichlorophenolxyacetic acid (2,4-D) at different time points of 2, 6, 12, 24, and 72 h. Using online tools at NCBI, DATF, and DRTF websites, 80 auxin-related genes changed at least at one time point were detected with the 41 up-regulated and 29 down-regulated during the whole period. The remained 10 genes showed up-regulation or down-regulation at different time points. Up-regulated genes were more than down-regulated genes, especially at 24 h and 72 h points. These genes were involved in several biological processes, such as transportation, response, induction, synthesis, and degeneration of auxin. Using semi-quantative RT-PCR technique, the expression changes of seven genes, coded as BG906698, BQ281752, CD454626, CD864552, CD938626, BJ233383, and AY543630, were confirmed to be consistent with the result by microarray. Gene of plasma membrane (PM) H⁺-ATPase (GenBank accession number: AY543630) expressed at a high level at the 2 h time point, and then decreased gradually till the minimum at the 24 h time point. This result implied that PM H⁺-ATPase gene might be a trigger in dedifferentiation of mature embryo in wheat.

Microsatellite Mapping of Powdery Mildew Resistance Gene in Wheat Landrace Xiaobaidong

XUE Fei,ZHAI Wen-Wen,DUAN Xia-Yu,ZHOU Yi-Lin,JI Wan-Quan

Acta Agron Sin. 2009, 35(10):  1806-1811.  doi:10.3724/SP.J.1006.2009.01806

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Xiaobaidong, a Chinese landrace, is proved to be resistant to powdery mildew (Blumeria graminis DC f. sp. tritici) with wide spectrum of resistance to pathogen races, especially effective to current prevalent races. This variety shows a response pattern that is different from the variety possessing documented Pm genes after inoculation with 10 isolates of different virulence. Genetic analysis indicated that a single recessive gene was responsible for the powdery mildew resistance at seedling stage. To detect the adjacent molecular markers of the gene, we constructed two mapping populations derived from the reciprocal crosses between Xiaobaidong and a susceptible cultivar Chancellor with 125 and 107 individuals of the F₂ generation. Seedlings of F₁ and F₂ plants were inoculated with isolate E09 of B. graminis DC f. sp. tritici for powdery mildew resistance identification. Using resistant and susceptible bulks, we identified the resistance gene mlxbd between simple sequence repeat (SSR) markers Xgwm577 and Xgwm1267 on chromosome 7. The gene mlxbd was located on the distal bin of chromosome 7BL-10 (Bin 0.78-1.00) by using the Chinese Spring nullisomic-tetrasomics, ditelosomics, and deletion lines. To verify the efficiency of the SSR markers that might be used in molecular marker-assistant selection (MAS), genome DNA of 23 wheat varieties with definite resistance genes were amplifiedby Xgwm577, the proximal marker of mlxbd. The result showed that Xgwm577 was able to differentiate resistance genes originated from Chinese varieties with Pm5a and Pm5b except PmH, however, resistance genes near Pm5 locus were failed to be identified with Xgwm577. This indicates that Xgwm577 should be combined with other markers in MAS.

QTL Mapping for Yield Traits in Upland Cotton(Gossypium hirsutum L.)

QIN Yong-Sheng,LIU Ren-Zhong,MEI Hong-Xian,ZHANG Tian-Zhen,GUO Wang-Zhen

Acta Agron Sin. 2009, 35(10):  1812-1821.  doi:10.3724/SP.J.1006.2009.01812

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As a major source of fiber and the world’s second-most important oil-seed crop after soybean, cotton plays an important role in the global economy. With the development of textile technology and social demand, it is urgent to breed and plant cotton varieties with high yield and super fiber quality. If the marker tightly linked with major gene controlling desired traits was identified, the efficiency of selection for agronomic traits might improve greatly. So far, high-identity genetic linkage map derived from upland cotton cultivars was lack because of their narrow genetic basis. Increasing the map density and tagging QTLs related with agronomic traits in Upland cotton will accelerate the process of marker assisted selection (MAS) breeding. CRI28 and XZM 2 are two cotton hybrids with high heterosis, which were bred by crossing CRI12, 4133, and 8891, respectively, with CRI12 as mutual parent. In this paper, two F₂ mapping populations were respectively assembled by using the parents of CRI28 hybrid (CRI12 and 4133, their corresponding population named as Pop1) and the parents of XZM 2 hybrid (CRI12 and 8891, their corresponding population named as Pop2), further, a joinmap linkage map which contained 245 loci and covered 1 847.81 cM was integrated with 27 mutual polymorphic loci in the two mapping populations by JOINMAP 3.0 software, after screening about 6 000 SSR primers. By the composite interval mapping method (CIM), the QTLs for eight yield-related traits in F₂ and F₃ populations were mapped. Of them, 43 QTLs were identified in the three environments by separating analysis and 16 QTLs by the joint analysis in Pop1; similarly, 66 and 20 QTLs in Pop2 respectively. Some QTLs on chromosome A3, D8, and D9, and some stable QTLs not influenced by environment were also detected. Twelve QTLs for eight traits could be found simultaneously in the two populations, and additive QTLs for fruit branches per plant, lint percentage and seed index all were offered by CRI12, suggesting that the value of CRI12 in breeding is mostly contributed by increasing the offspring’s bolls. These results will provide very important information in Upland cotton breeding for yield by MAS.

Cloning and Expression Profiling of Gibberellin Insensitive Dwarf GID1 Homologous Genes from Cotton

DONG Jing,YIN Meng-Hui,YANG Fan,ZHAO Juan,QIN Shan,HOU Lei,LUO Ming,PEI Yan

Acta Agron Sin. 2009, 35(10):  1822-1830.  doi:10.3724/SP.J.1006.2009.01822

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Gibberellins (GA) play an important role in regulating cotton fiber development. As the GA receptor, GID1 is an essential component of GA signaling pathway. To advance our understanding of GA’s functions in cotton development (especially fiber development), we cloned six cotton GID1 homologous genes (GhGID1-1–6, accession No. are FJ790125–FJ790130) by searching EST sequences, extending the flanking sequences by Y-shaped adaptor dependent extension (YADE) method and amplifying the full-length cDNA by 3'-RACE. Multiple sequence alignment indicated that the deduced GhGID1-1–6proteins shared high sequence identity with GID1s from other species and contained multiple binding sites with GA and DELLA protein as well as HGG and GXSXG domains conserved in hormone-sensitive lipase (HSL) family. Three amino acid residuals (G¹⁶⁹, G¹⁹⁶, and R²⁵¹) with the rice GID1 function were also conserved in cotton GID1s. In the phylogenetic tree of GhGID1-1–6 and other GID1 proteins with known function, GhGID1-1–6 were clustered with 3 Arabidopsis GID1 proteins, and distantly related to GID1s from rice and S. moellendorffii. Quantitative RT-PCR was employed to detect the expression levels of GhGID1-1–6 in various organs and tissues. The transcripts of GhGID1-1–6 could be detected in all the organs and tissues investigated (including roots, hypocotyls, leaves, petals, anthers, ovules and fibers), although with various levels. GhGID1-1 and GhGID1-2 expressed mainly in floral organs, while GhGID1-4 expressed preferentially in fiber and root. Exogenous GA added in medium led to alteration of gene expression level in the in vitro cultivated ovules, and the expressions of GhGID1-1 and GhGID1-2 were obviously inhibited by GA. These results suggested that GhGID1-1–6 might encode biologically functional GID1 homologues involved in GA signaling in cotton. To further clarify GID1 genes related to fiber development, we compared GhGID1s’ expression in developing ovule (from 6 to 18 days post anthesis, DPA) and the attached fibers by quantitative RT-PCR. GhGID1-1 was predominantly expressed in ovules, and its expression level reached the climax at 10 DPA, and kept the high level at 14 DPA and 18 DPA, while in fibers, GhGID1-4 was the main GID1 homologue expressed, and its highest-level expression occurred at 6 DPA, and declined to a very low level at 14 DPA and 18 DPA.This result strongly suggested that these are relatively independent GA signaling systems in ovules and attached fibers.

Establishment and Application of Multiples-PCR for Wheat Glutenin Subunits Relative to Superior End-Use Quality

ZHENG Han,CHEN Jing,REN Yan,YU Mao-Qun,FU Ti-Hua

Acta Agron Sin. 2009, 35(10):  1831-1835.  doi:10.3724/SP.J.1006.2009.01831

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Wheat processing qualities are largely dependent on the number and composition of high-molecular-weight glutenin subunits (HMW-GS). It has been demonstrated that Ax1/Ax2*, Dx5, and overexpressed Bx7 (Bx7^OE) are normally associated with superior end-use quality, especially dough strength. So far, Bx7^OE allele has not been exploited widely in China wheat breeding program. With the optimization of reaction components and amplification condition, three pairs of PCR primers targeting for Ax1/Ax2*, Bx7^OE, and Dx5 genes were used to establish a multiplex PCR for the purpose of enhanced molecular marker-assisted breeding with good reliability and low cost. The validation with wheat cultivars (lines) carrying known genes displayed that the developed multiplex PCR permitted the discrimination of these major HMW-GS in a single PCR reaction and agarose gel assay. A total of 89 wheat cultivars and advanced lines from Tibet were tested by the multiplex PCR-based assay. The results showed that each frequency of Ax1/Ax2* and Dx5 was 12.4%, and only 10.1% of the accessions possessed both Ax1/Ax2*and Dx5, whereas no accessions carried Bx7^OE. Introduction of HMW-GS relative to good gluten quality must enhance the improvement of wheat quality in this region.

Map Integration of QTLs for Grain Yield and its Related Traits in Maize

WANG Bang-Tai,WU Jian-Yu,Ding Jun-Qiang,XI Zhang-Ying

Acta Agron Sin. 2009, 35(10):  1836-1843.  doi:10.3724/SP.J.1006.2009.01836

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Identification and fine mapping of quantitative trait loci (QTLs) for grain yield and its related traits in maize are very important for molecular breeding by design. In the past few decades, a wealth of QTLs mapping data for grain yield and its related traits in maize has been produced using molecular marker approaches. In order to unlock the full potential of the information contained in these independent experiments, four hundreds QTLs for grain yield and its related traits in maize, collected from different publications, were used to construct new QTL integrated map and consensus map using bioinformatics and meta-analysis methods with IBM2 2008 neighbors as reference. It showed that these QTLs were distributed on all 10 chromosomes unevenly, with the most on chromosome 1 and the least on chromosome 10. QTLs for ear length, cob diameter, kernel number per row, kernel weight and grain yield were mainly distributed on chromosome 1, while QTLs for ear row number on chromosome 9. Ninety-six “Consensus” QTLs for grain yield and its related traits were estimated, including 43 “Consensus” QTLs for kernel weight, 32 “Consensus” QTLs for grain yield, and 8, 5, 4, 3, 1 “Consensus” QTLs for ear diameter, ear row number, cob diameter, kernel number per row, ear length, respectively. QTLs with similar phenotype were clustered on the same or near locations. These results provide a good basis for studying genetic mechanism and molecular marker- assisted selection for grain yield and its related traits in maize

Molecular Identification of Phytophthora Resistance Gene in Soybean Cultivar Yudou25

FAN Ai-Ying,WANG Xiao-Ming,FANG Xiao-Ping,WU Xiao-Fei,ZHU Zhen-Dong

Acta Agron Sin. 2009, 35(10):  1844-1850.  doi:10.3724/SP.J.1006.2009.01844

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Phytophthora root rot, caused by Phytophthora sojae, is a destructive disease on soybean. Use of resistant soybean cultivars is the most economical and effective method for controlling the disease. Up to now, nine loci for the resistance with 15 genes have been identified in soybean. However, only a few genes, such as Rps1c and Rps1k,were effectively resistant to populations of P. sojae in China, so mining new resistance genes is necessary greatly for the disease control. Soybean cv. Yudou 25 has broad spectrum resistance to P. sojae, and is an elite resistance source for Phytophthora root rot of soybean. To effectively utilize the cultivar in resistance breeding, in the present study, we identified and tagged the Phytophthora resistance gene in the cultivar by using SSR markers and bulked segregation analysis (BSA). Two F_2:3 populations were developed for resistance genetic analysis and resistance gene mapping. Using hypocotyls inoculation technique at the seedling stage in the glasshouse, the reaction to P. sojae isolate PSMC1 (virulence type 1b, 1d, 3a, 3b, 3c, 4, 5, 6, 7) in 82 and 98 F_2:3 families derived from two crosses of Yudou 21×Yudou 25 and Zaoshu 18×Yudou 25, respectively, were identified. The segregation ratio in both populations fit into 1:2:1 for homozygous resistant, segregating and homozygous susceptible, showing that the cultivar resistance to Phytophthora root rot is controlled by a dominant single gene, with the temporary name of RpsYD25. On the basis of linkage analysis with SSR markers, RpsYD25 was located on soybean molecular linkage group (MLG) N in both populations. Five SSR markers were associated with RpsYD25 in an order of Sat_208-Satt530-RpsYD25-Sat_084-Satt125-Sat_236 in F_2:3 population from the cross of Yudou 21×Yudou 25, RpsYD25 was flanked by Satt530 and Sat_084 with a distance of 6.3 and 7.7 cM, respectively. Five SSR markers were linked to RpsYD25 in an order of Satt125-RpsYD25-Sat_275-Sat_266-Satt660-GMABAB in F_2:3 population from the cross of Zaoshu 18×Yudou 25, RpsYD25 was flanked by Satt125 and Sat_275 with a distance of 7.9 and 7.8 cM, respectively. Because RpsYD25 was mapped on MLG N near to Rps1 locus, the genetic relationship of RpsYD25 and Rps1 was detected by using the selected SSR markers contained in the Rps1k allele sequence. A SSR marker Rps1k6 in Rps1k allele was found to be linked to RpsYD25 with a genetic distance of 19.4 cM in F_2:3 population from the crosses Yudou 21×Yudou 25. Therefore, the Phytophthora resistance gene RpsYD25 in cv. Yudou 25 might be a novel allele at Rps1 locus, or a novel gene.

A New Rice Cultivar“Wulingjing1” with Resistance to Rice Stripe Virus Bred by Marker Assisted Selection

PAN Xue-Biao,CHEN Zong-Xiang,ZUO Shi-Min,ZHANG Ya-Fang,WU Xu-Jiang,MA Ning

Acta Agron Sin. 2009, 35(10):  1851-1857.  doi:10.3724/SP.J.1006.2009.01851

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Rice stripe virus disease is one of the most serious diseases in rice production area of Jiangsu province since 2002. A commercial rice cultivar with excellent food taste-quality, Wuyujing 3, was highly susceptible to the disease and was improved through the MAS strategy in a backcross project with “Zhendao 88” as donor parent of resistance gene Stvb-i. In each backcross generation, we selected the progenies with Stvb-i based on the closely flanked markers. In advanced backcross generations, the marker’s detection for genetic background of selected plants was performed, along with the estimate by the eye for the similarity of selected plants to recurrent parent in synthetic agronomic traits. As a result, a new cultivar, Wulingjing 1, was fleetingly bred in BC₃F₄ generation, whose resistance to the disease was significantly increased and main characters especially on excellent taste-quality were consistent with those of the recurrent parent Wuyujing 3. In resistance identification tests with multiple locations organized by Seed Management Station of Jiangsu Province, the average percent of affected plants of the new cultivar was only 4.4% and significantly lower than that of recurrent parent (53.2%).

Genetic Diversity of Adzuki Bean Germplasm Resources Revealed by SSR Markers

WANG Li-Xia,CHENG Xu-Zhen,WANG Su-Hua,LIANG Hui,Zhao Dan,XU Ning

Acta Agron Sin. 2009, 35(10):  1858-1865.  doi:10.3724/SP.J.1006.2009.01858

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Adzuki bean, originated from China, is either one of traditional exports or an important crop for agricultural structure adjustment. As the traditional varieties can not meet the demands of modern markets and the genetic bases of the cultivars have been narrow, it is necessary to develop new breeding materials or novel germplasm, to improve varieties of adzuki bean. China has a collection of over 5 000 adzuki bean resources, which is lacking in study and use in breeding. In the paper, genetic diversity of 158 accessions of cultivated and 18 wild types of adzuki beans were analyzed by using 24 pairs of SSR primers. The results showed that 18 pairs of SSR primers were detected to be polymorphic in cultivated adzuki beans and 21 in wild adzuki beans. The average number of allele (NA) per SSR locus for cultivated and wild adzuki beans was 3.0 and 2.6, respectively. The number of accessions used for cultivated and wild adzuki beans differed much, and the NA and genetic diversity index (polymorphic information content, PIC value) showed a higher genetic variation in wild types than in cultivated ones. Both cluster and principal coordinate analysis (PCO analysis) suggested that there was a high genetic differentiation between cultivated and wild adzuki beans, while cultivated adzuki beans from different provinces of China could not be distinguished from each other clearly, suggesting that there is a high rate of the gene exchange among local adzuki beans. The genetic backgrounds of the breeding cultivars revealed by SSRs are almost agreeable with their putative pedigree, indicating that these SSRs can be used to identify the genetic relationships of them, and used in marker-assist-selection (MAS) in breeding. The results also revealed that the adzuki bean germplasms occurred more than once with the same name in the National Germplasm Conservation Centre were different from each other in genetic background and so valuable to be conserved and used individually. The present study not only provides some informations in adzuki bean breeding, but can accelerate the application of SSR markers in diversity analysis, gene tagging and identification of new cultivars.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY

Effects of Nitrogen Nutrition on Grain Quality in Upland Rice Zhonghan 3 and Paddy Rice Yangjing 9538 under Different Cultivation Methods

ZHANG Ya-Jie,CHEN Ying-Ying,YAN Guo-Jun,DU Bin,ZHOU Yu-Ran,YANG Jian-Chang

Acta Agron Sin. 2009, 35(10):  1866-1874.  doi:10.3724/SP.J.1006.2009.01866

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Upland rice and dry-cultivated paddy rice have been attracted more and more attention because of limited water resources in China. Researches on interaction between water and nitrogen supplies for crop resistance to drought stress has become a hot topic in crop physiology. However, the information linking to the effect of nitrogen nutrition on grain quality of upland rice and paddy rice under different cultivation methods is unavailable. The objective of this study was to evaluate the difference between upland rice and paddy rice and interaction between cultivation methods and N levels. One upland rice cultivar Zhonghan 3 (japonica) and one paddy rice cultivar Yangjing 9538 (japonica) were grown under either moist cultivation (MC, control) or bare dry-cultivation (DC) conditions, with three N levels, low amount of nitrogen (LN, 100 kg ha^-1), normal amount of nitrogen (NN, 100 kg ha^-1), and high amount of nitrogen (HN, 300 kg ha^-1). The results showed that, compared with NN, the grain yield under HN was lower for both upland and paddy rice under DC and for paddy rice under MC, whereas it was higher for upland rice under MC. Under both DC and MC，the percentage of chalky grains and the chalkiness of upland rice were increased under NN and reduced under HN. With the increase in N levels, amylose content was reduced and protein content was increased for both upland and paddy rice. However, the percentage of chalky grains and the chalkiness of paddy rice was decreased under DC and increased under MC. Breakdown viscosity was the highest and setback viscosity was the lowest for upland rice under NN and for paddy rice under LN. The bare dry cultivation can improve appearance quality and nutrient quality for upland rice. There was no significant difference in other rice quality indices between upland rice and paddy rice under DC. Compared with paddy rice, upland rice showed better nutrient quality and poor appearance quality and cooking quality. The correlation coefficient between cooking and nutrient quality and leaf nitrogen content was smaller for upland rice than for paddy rice. The results suggest that the response to cultivation methods and nitrogen levels varies largely between upland rice and paddy rice. The approaches to increase grain yield and grain quality for both paddy and upland rice were discussed.

Effects of  Plant Density on Grain Number and Grain Weight at Different Spikelets and Grain Positions in Winter Wheat Cultivars

QU Hui-Juan,LI Jin-Cai,SHEN Xue-Shan,LI Ru-Yi,WEI Feng-Zhen,ZHANG Yi

Acta Agron Sin. 2009, 35(10):  1875-1883.  doi:10.3724/SP.J.1006.2009.01875

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Seed-setting traits and grain weight are influenced by either genetic factor or cultivation environment in wheat (Triticum aestivum L.), such as plant density. Due to the unbalanced development of a spike, grain number and grain weight at different spikelets and grain positions are largely different. Although their distributions have been described in earlier reports, the effect of plant density is not clear. This study focused on the effect of plant density on the grain number and grain weight of two wheat cultivars (Lankao Aizao 8 of large-spike type and Zhoumai 18 of medium-spike type) at different spikelets and grain positions. The grain number, spikelet weight, and grain weight with the spikelets positions from the bottom to the top increased first and then decreased. The differences of seed-setting traits between two cultivars were significant. Meanwhile, seed-setting traits of main sipke were supeirior than that of tillering spike. Under low sowing density, the seed setting characteristics and grain weight in Lankao Aizao 8 were more superior than those in Zhoumai 18, and those of main stem spike were more superior than those of tillering spike. The effects of spikelet and grain positions on the weight of individual grain varied with the grain number of spikelet. For the basal and middle spikelets, the grain weight was higher at second grain position than at the first one, whereas it was inverse for the upper and top spikelets. The grain weight was lower in the third grain position than in the first and second grain position, and the least in the fourth grain position. In conclusion, appropriate plant density is important for high yield of different spike-type cultivars, and in wheat production we should properly increase the proportion of tillering spike on the basis of ensuring main stem spike number; increase grain weight on the basis of ensuring fertile grain number especially for basal spikelet; and develop the potential of grain weight in the third and fourth grain positions on the basis of ensuring that in the first and second grain positions of spikelet.

Effects of Irrigation Stage and Amount on Water Consumption Characteristics,Flang Leaf Photosynthesis,and Grain Yield in Wheat

MENG Wei-Wei,ZHANG Yong-Li,MA Xing-Hua,SHI Yu,YU Zhen-Wen

Acta Agron Sin. 2009, 35(10):  1884-1892.  doi:10.3724/SP.J.1006.2009.01884

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Shortage of water resource has become one of the major factors limiting wheat (Triticum aestivum L.) production in North China. Irrigation plays an important role to obtain high grain yield. Currently, studies on water consumption characteristics and the physiological basis of yield formation in wheat are seldom reported. In this study, three wide planted cultivars, Jimai 20, Taishan 23, and Taishan 22 were used in two independent experiments in 2004–2005 and 2005–2006 growing seasons. There were four irrigation treatments in both experiments, i.e., no irrigation (W0), soil water plus one irrigation at jointing stage (W1), soil water plus two irrigations at jointing and anthesis stage (W2), soil water plus three irrigations at jointing, anthesis, and grain-filling stage (W3). Each irrigation supplied water of 60 mm. In the 2004–2005 growing season, the highest grain yields of Jimai 20 and Taishan 23 were obtained in W2 treatment with water consumptions of 429.8 and 453.0 mm, respectively, which were higher than those of W1 treatment (402.9 and 416.6 mm). However, the irrigation efficiency of W2 treatment was lower than that of W1 treatment. The water use efficiencies (WUE) of W1 and W2 treatments were both higher than those of other treatments in the two cultivars. In Jimai 20, there was no significant difference between W1 and W2 treatments, whereas in Taishan 23, W1 had higher WUE than W2. In the 2005–2006 growing season, Jimai 20 yield the highest in W1 treatment, and Taishan 22 in W2 treatment, with water consumptions of 451.3 and 459.2 mm, respectively. The highest and the lowest WUE were in W0 and W3 treatments in both Jimai 20 and Taishan 22, respectively. In Jimai 20, the WUE was higher in W1 than in W2, whereas there was no significant difference between W1 and W2 in Taishan 22. With the increase of irrigation amount, the percentage of soil water supply amount and precipitation to total water consumption amount decreased and the percentage of irrigation amount to total water consumption amount increased. Compared with W1, W2, and W3 treatments, W0 treatment had lower flag leaf photosynthetic rate and sucrose phosphate synthase activity at medium and late grain-filling, but at early grain-filling, there was no significant difference between W0 and W1, W2, W3, and the transfer of sucrose from flag leaf to grain was more favorable in W0 than in W1, W2, and W3, thus, Jimai 20 obtained the highest grain weight in W0 treatment. In wheat production under conditions similar to this study, the optimal irrigation regimes are suggested as 60–120 mm for Jinmai 20 and Taishan 23 at jointing or at jointing and anthesis stages without irrigating base water, and 60 mm for Jinmai 20 at jointing or 60–120 mm for Taishan 22 at jointing or at jointing and anthesis stages with irrigating base water 60 mm .

Effect of Nitrogen and 6-BA on Development of Tillering Bud and Its Physiological Mechanism

LIU Yang,WANG Qiang-Sheng,DING Yan-Feng,LIU Zheng-Hui,LI Gang-Hua,WANG Shao-Hua

Acta Agron Sin. 2009, 35(10):  1893-1899.  doi:10.3724/SP.J.1006.2009.01893

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Tiller number is a major factor affecting rice yield, nitrogen and cytokinin have substantial regulative effect on tillering of rice. However, little is known on the mechanism of nitrogen and cytokinin regulating tillering bud growth and on the relationship of them during this process. In this study, two rice cultivars (Yangdao 6 and Nanjing 44) were used to investigate the effect of nitrogen and cytokinin on the regulation in tillering bud growth and the changes of nitrogen, carbon and cytokinins in plant during this process. The results showed that both nitrogen and cytokinin promoted the germination of tillering bud, but there was a significant difference in the effect of them on the subsequent outgrowth, nitrogen promoted both germination and outgrowth of tillering bud, however, cytokinin only promoted tillering bud’s germination. Both nitrogen and 6-BA significantly increased cytokinin (Z+ZR and iP+iPR) content in tillering node and promoted activity of nitrate reductase (NRA) in leaf and root, but 6-BA couldn't significantly increase the content of soluble protein, nitrogen and non structural carbohydrate (NSC) in plant. In conclusion, nitrogen and 6-BA may regulate tiller production by two approaches, one is the promotion of endogenous hormones balance and the other is the controlling of carbon and nitrogen metabolism.

Characteristics of Source-Sink Related Parameters in Conventional Indica Rice Cultivars with Different Types of Sink Potential

DONG Gui-Chun,TIAN Hao,DONG Yan-Ping,ZHANG Biao,LI Jin-Qian,YU Xiao-Feng,

Acta Agron Sin. 2009, 35(10):  1900-1908.  doi:10.3724/SP.J.1006.2009.01900

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Eighty-eight and one hundred and twenty-two conventional indica rice cultivars were solution-cultured in 2001 and 2002, respectively. Leaf area index (LAI), dry matter weight (including root, culm and sheath, leaves, panicle), nitrogen content in different organs, yield and its components were measured respectively. The tested rice cultivars were classified into 6 types (i.e. A, B, C, D, E and F, A was the lowest, and F was the highest) based on their sink potential level by the MinSSw method, to study their difference in source and sink. Results were as follows: (1) Difference in sink potential (SP) of the cultivars used in this study was very large, the averages of SP in types of A, B, C, D, E, F were 426.37, 642.53,770.96, 903.73, 1 064.32, 1 213.90 g m^-2 in 2001, and 359.36, 574.11, 764.98, 962.43, 1 200.11, 1 455.59 g m^-2in 2002, respectively. (2) SP was significantly influenced by growth duration and SP per day, and the effect of SP per day on SP was more important than that of growth duration. (3) LAI at heading and maturing stage, green leaf weight at heading and maturing stage, leaf weight per unit leaf area in large SP type of indica rice were higher than those in small SP type, while the decline proportion of LAI during grain filling period. (4) Varieties with large SP showed higher net assimilation rate and higher values in grain yield per unit leaf area, SP per unit leaf area, SP per unit dry matter weight and SP per unit N at heading stage. (5)Multiple stepwise regression analysis indicated that SP was significantly influenced by green leaf weight at heading stage, SP per unit leaf area, SP per unit N, SP per unit dry matter weight, net assimilation rate during grain filling period and leaf weight per unit leaf area, with the determination coefficient ranging from 0.953 to 0.963. Path analysis showed that effects of green leaf weight at heading stage and SP per unit leaf area on SP were larger than those of SP per unit N, and SP per unit dry matter weight, net assimilation rate during grain filling period, and leaf weight per unit leaf area.

Forms and Accumulation of Aluminum in Tea Plant(Camellia sinensis)

SUN Ting,LIU Peng,ZHENG Ren-Wei,XIE Zhong-Lei,LUO Hong

Acta Agron Sin. 2009, 35(10):  1909-1915.  doi:10.3724/SP.J.1006.2009.01909

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Aluminum toxicity is a major limiting factor affecting yield and quality of crops in acid soil. Camellia sinensis is an Al-accumulating plant grown healthily even in strong acid soil with high aluminum content. The study of the forms and accumulation of aluminum will be helpful to reveal the Al-toleranting mechanism of tea plant. In this paper, the content of Al³⁺ in tea plants was determined after Camellia sinensis wastreated for 50 d with different concentrations of Al³⁺ in culture solution. Besides, the forms of Al were analyzed in the roots, stems and leaves of tea plants by the nondestructive ²⁷Al NMR with high accuracy. The old leaves of tea plant were the major Al-accumulating organ, whereas, roots would take the position of it and accumulate more aluminum when the plant was treated with 2 mmol L^-1 Al³⁺. ²⁷Al NMR test showed that the dissoluble aluminum in tea root, stem and leaf existed mainly in the form of [AlO₄Al₁₂(OH)₂₄(H₂O)₁₂]⁷⁺(Al₁₃), whose chemical shift was 63×10^-6, and it was the first discovery in tea plant. Besides of Al₁₃, there were two weak absorption peaks of –0.38×10^-6 and –0.17×10^-6 in three organs tested, which wereundetected aluminum complexes. Other three complexes appeared in old leaf when treated by 5 mmol L^-1 Al³⁺, which were Al-oxalate (1:1), Al-oxalate (1:2), and Al-phosphate, and the chemical shifts were respectively 6.4×10^-6, 12.7×10^-6 and –6.2×10^-6. In summary, most of Al³⁺in tea plant exist in chelate complexes or organic matter, which can be considered as one of Al-accumulating and Al-tolerance mechanism of tea plant.

Effects of Drought Stress on Activity of Photosystems in Leaves of Maize at Grain Filling Stage

LI Geng,GAO Hui-Yuan,ZHAO Bin,DONG Shu-Ting,ZHANG Ji-Wang,YANG Ji-Shun,

Acta Agron Sin. 2009, 35(10):  1916-1922.  doi:10.3724/SP.J.1006.2009.01916

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At grain filling stage, the effects of drought stress on photosynthetic acivities of photosystem I (PS I) and photosystem II (PS II) in leaves of maize (Zhengdan 21, a cultivar with high starch content) were studied by simultaneously analyzing chlorophyll a fluorescence transient and light absorbance at 820 nm. The results, obtained from two years experiments, demonstrated that the drought stress significantly reduced photosynthesis (P<0.05) and grain yield (P<0.05) of the maize. The K and J steps at fluorescence transient were increased by drought stress, which indicated the inhibition of oxygen- evolving complex (OEC) and electron transport chain after Q_A in PS II. The acceptor side of PSII was damaged more severely than the donor side of PSII. Furthermore, the maximal oxidation-reduction activity of PS I (ΔI/I_o) was also significantly decreased by the drought stress, which inhibited the photosynthetic electron tranport from the PS II to PS I, destructing the coordination between PS I and PS II. We suggest that the inhibition of PS I and PS II and the destruction of the coordination between PS I and PS II by the drought stress is one of the main reasons to cause the decrease in photosyntheis and grain yield of maize.

Relationship Between Dry Matter Per Plant in First Cutting and the Yield-Related Traits in Twelve Alfalfa Varieties Introduced Overseas

LONG Ming-Xiu,GAO Jing-Hui,GAO Yang,HU Tian-Ming,SHI Jun-Tong,WU Zhen,

Acta Agron Sin. 2009, 35(10):  1923-1929.  doi:10.3724/SP.J.1006.2009.01923

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Yield of alfalfa (Medicago sativa L.) is closely related to its plant characteristics and the yield in 1st cutting contributes the majority to the annual harvest with the dry matter per plant (DMPP) determining the total community yield. The relationship between dry matter per plant in 1st cutting and yield-related traits (plant height, branch characteristics and leaf to stem ratio) of 12 introduced alfalfa varieties were studied in three continuous years (the 3–5 growth years) under field condition. There was a significant positive correlation between dry matter per plant and sub-branch number (SBN) (r = 0.689), and between DMPP and internode number of sub-branch (SBIN) (r = 0.526). The 1stprincipal component was composed of the branch number (BN), SBN, branch internode number (BIN) and SBIN, which accounted for 58.05 percent of total variance. This suggests that SBN and SBIN may be considered as the main objective characteristics for alfalfa breeding selection. The plant height (PH) had no significant correlation with DMPP, but a significant negative correlation with BN and BIN (r = –0.650 and –0.637, respectively), indicating that the control of the plant height of alfalfa by filed management appropriately is beneficial to the yield. The cluster analysis for DMPP showed that the 3-year average SBN in 1st cutting for group 1 varieties (the group with the highest DMPP including Pondus and WL-414) was significantly higher than other two groups. The SBIN had no significant difference between group 1 and group 2, but showed significant difference between group 1 and group 3 (P<0.05). The leaf to stem ratio (LSR) of group 1 was 6.05% higher than that of group 2 and 10.91% higher than that of group 3, and the difference was significant between group 1 and group 3 (P<0.05). Among the varieties, Pondus and WL-414 had higher breeding value in yield and palatability than other varieties.

RESEARCH ACTIVITIES

Comparison of Photosynthetic Characteristics of Leaves in Pop Corn，Sweet Corn and Glutinous Corn during the Late Growing Period

ZUO Zhen-Peng,SUN Qing-Quan,DONG Lu-Hao,WANG Jing,MA Deng-Chao,DONG Shu-Ting

Acta Agron Sin. 2009, 35(10):  1930-1935.  doi:10.3724/SP.J.1006.2009.01930

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The differences were studied among pop corn, sweet corn and glutinous cornin photosyntheticcharacteristics of leaves in different positions of the plants during the late growing period in the field conditions. The results showed thatdifferences of photosynthetic characteristics of leaves were superior for glutinous corn than for the sweet corn and pop corn. Moreover all the indices of photosynthesis rate (P_n), content of chlorophyll (Chl), transpiration rate (T_r) and stomatal conductance (G_s) were glutinous corn>sweet corn>pop corn. The photosynthesis rate of mid-leaves was glutinous corn>sweet corn>pop corn, whereas the difference of photosynthesis among lower leaves in plant was not significant. The chlorophyll content (Chl) of top and middle leaves was pop corn>sweet corn>glutinouscorn. Correlation analysis showed that it was significant at 5% or 1% probability levels in the different growing periods between P_n and Chl, T_r, G_s. At the same time, the photosynthetic characters of leaves in the top and middle positions of plant excelled those in the low leaves for all the three corn types. The P_n, Chl, duration for high P_n and T_r of glutinouscorn were superior to pop corn and sweet corn; attenuation time for leaf photosynthetic capability of pop corn and sweet corn was earlier than that for glutinous corn.

Genetic Diversity Analysis of Black Sesame(Sesamum indicum DC) Core Collection of China Using SRAP Markers

CHE Zhuo,ZHANG Yan-Xin,SUN Jian,ZHANG Xiu-Rong,SHANG Xun-Wu,WANG Hua-Jun

Acta Agron Sin. 2009, 35(10):  1936-1941.  doi:10.3724/SP.J.1006.2009.01936

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China has a long history and rich germplasm resources for growing black sesame. Black sesame is well used because of its nutrition superior to black rice and black soybean. However, Research on black sesame is relatively less than that on other crops. In the paper, genetic diversity of black sesame core collection of China was analyzed using SRAP (sequence-related amplified polymorphism) markers. The results showed that 13 SRAP primer combinations were employed to evaluate the genetic diversity of 100 black sesame accessions, a total of 182 amplified fragments were detected and 126 of them were polymorphic, the polymorphism percentage was 69.2%, the number of amplified fragments and polymorphic fragments of each primer combination were 14.0 and 9.7, respectively. The 100 accessions were grouped into five clusters at genetic similarity of 0.68, indicating the genetic diversity of these accessions was abundant. As regards geographic regions, Shannon's information index of black sesame accessions in south China (0.3557) was the highest，the followings were that in central regions of China (0.3415) and north China (0.2986). The characteristics of genetic diversity of black sesame core collection of China were fully revealed in this study, which provided the theoretical foundations for further exploring, collecting and introducing black sesame germplasm as well as mining and utilizing excellent black sesame germplasm in the future.

In silico Mapping and Structure Analysis of Key Enzyme Genesis in Fatty Acid Synthesis of Soybean

SONG Wan-Kun,ZHU Ming-Xi,ZHAO Yang-Lin,WANG Jing,LI Wen-Fu,LIU Chun-Yan

Acta Agron Sin. 2009, 35(10):  1942-1947.  doi:10.3724/SP.J.1006.2009.01942

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The 12 genes of key enzyme such as acyl-CoA carboxylase and fatty acid synthase in fatty acid synthesis were mapped on the newest genetic map integrating from physical map and genetic map, and the gene structure was analyzed. They were mapped on nine linkage groups, including A2, B2, C2, D1b, D2, G, I, L, M, and the flank markers of the gene on the linkage groups were obtained. At the same time, we compared the sequence information of cDNA with gDNA to get the structure information of the 12 genes, with the number of intron from 0 to 30. FAD2-1, FAD2-2, FAD2-3, and FatB were all single extron gene, but there were six introns in KASI, twelve introns in KASII, two introns in SACPD, and seven introns in FAD3. So the corresponding markers obtained from the mapping are available for molecular assisted selection, while the structure information can be better used in gene function analysis.