http://www.molplantbreed.org/ Table of contents of Molecular Plant Breeding Zhu Yongsheng (1,2,3,4), Wang Yingheng (1,2,3,4), Tu Shihang (2,3,4), Wu Fangxi (1,2,3,4), Cai Qiuhua( 1,2,3,4), Zheng Jiatuan (1,2,3,4), Zhang Jianfu (1,2,3,4*), Xie Hua'an (1,2,3,4*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1254 The photosynthetic rate of different restore lines and their combinations was measured by Li6400 in different growth period. The results indicated that the photosynthetic rate was different for different varieties in different growth period. At the heading period, the maxium photosynthetic rate was 19.2 ¦Ìmol¡¤m^-2¡¤s^-1 for the variety Shanyou63 and the minium photosynthetic rate was 12.6 ¦Ìmol¡¤m^-2¡¤s^-1 for the variety Minghui86, but the difference of photosynthetic rate was notable among different varieties. At the grouting period, the maxium photosynthetic rate was 14.3 ¦Ìmol¡¤m^-2¡¤s^-1 for the variety ¢òyouhang1 and the minium photosynthetic rate was 12.1 ¦Ìmol¡¤m^-2¡¤s^-1 for the variety Hang1, but the difference of photosynthetic rate was not notable among different varieties. At the mature period, the difference of photosynthetic rate was not notable among different varieties, but the photosynthetic rate was different at the different time for the same variety, indicating that the intension of sunlight affected the photosynthetic rate for rice. The photosynthetic rate in different growth period was compared with among different varieties, indicating that the trend of photosynthetic rate was consistent among different varieties from the heading period to mature period. But the photosynthetic rate was larger in heading period than that in mature period. In the meanwhile, the components of yields indicated that the photosynthetic rate was related with the yields in some extend. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Li Jiayang http://www.molplantbreed.org/publish/abstract.asp?PaperID=1250 In 2008, two research groups headed by Dr. Qifa Zhang of Huazhong Agricultural University and Dr. Yaoguang Liu of South China Agricultural University published their latest research works about the indica-japonica hybrid sterility in rice in PNAS. They cloned S5 and Sa that are the major loci controlling female fertility with wide compatibility and male gamete abortion respectively in indica-japonica hybrids. They analyzed the molecular interactions between the alleles of the genes and proposed a triallelic system for S5 and a two-gene/three-component interaction model for Sa. These findings have greatly enhanced understanding of reproductive isolation with implication for the process of speciation, and will facilitate utilization of inter-subspecific heterosis in hybrid rice breeding. These are significant achievements in the research of plant hybrid sterility by Chinese scientists following the mechanistic elucidation of cytoplasm male sterility and fertility restoration in Boro¢òtype rice in 2006. Thu, 28 Jan 2009 13:40:45 GMT Invited Review Dong Jing, Zhang Yuping, Pan Qinghua, Li Shaoning, Jin Wanmei (*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1256 A cold-inducible transcription factor dehydration response element binding CBF1 from Arabidopsis thaliana was genetically transformed by using leaf-disc method via Agrobacterium-tumefaciens into the experimental material¡°Honeoye¡±strawberry. In 5 independent transgenic lines 80, 83, 94, 104 and 10-5-5, crossing treating experiments were carried out. Filial generations were screened by with 30 mg/L kanamycin, PCR and Southern blot. The foreign CBF1 gene could stability pass from parent to generations and the inheritance was not cytoplasmic heredity. In 83, 94, 104 and 10-5-5, the foreign CBF1 gene of F₁ progeny was integrated into the chromosome of strawberry generation and expressed abiding by genetic law, but line 80 was gene interaction. The foreign CBF1 gene of F₂ was complex. Because of obtaining progeny of single flower, genetic model was 3:1, 15:1, 35:1 segregation ratio. Meanwhile the foreign CBF1 gene of F₂ progeny existed gene interaction. The results showed that the foreign gene of transgenic strawberry had a complex inheritance. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Ouyang Dongmei (1,2*), Chen Shen( 1*), Jiang Junxi (2), Yang Jianyuan( 1), Zeng Liexian( 1), Zhu Xiaoyuan (1**) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1255 BTX, an indica rice mutant from Basmatic370, showed a broad spectrum resistance to Mangnaporthe grisea (Hebert) Barr. To identify and map the blast resistance genes in BTX, seven isolates 95-59a, 98-288a,00-193a, 05-20a, 05-135a, 06-138a and RB6 which are compatible to Lijiangxintuanheigu (LTH, very susceptible to rice blast) and incompatible to BTX, were used for inoculating a population of recombinant inbred lines (RILs) derived from the cross between BTX and LTH. The results showed that the segregation of the population to three isolates 95-59a, 98-288a and 05-20a fit to 3R:1S, indicating that their resistance are controlled by two resistance genes. The population segregated into 1R:1S against other four isolates 00-193a, 05-135a, 06-138a and RB6, suggesting that the resistance gene (s) detected in the RILs population is a single resistance gene. In present study, the isolate 00-193a with broader pathogenic spectrum and F2 population derived from the cross between LTH and BTX were used for resistance gene mapping. The F₂ population segregated into 3R:1S, indicating that the resistance gene detected in the F₂ population is a single dominant resistance gene, temporarily designed as Pibt(t). A total of 250 SSR markers selected from each rice chromosome equally were screened. The result showed that the marker RM224 located on chromosome 11 was linked to the resistance gene in the respective populations above. To confirm this result, four additionally polymorphic SSR markers, which located in the region beside RM224, were tested. The linkage analysis results showed that the genetic distances between markers RM27181, RM27272, RM224, RM27334 and RM37363 are 7.74 cM, 1.50 cM, 0.25 cM, 10.59 cM and 13.24 cM. Pibt(t) is mapped in an interval between RM224 and RM27334 with a genetic distance of 10.84 cM. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Mao Weiwei( 1,4), Gao Hongsheng (1), Bo Tianyue( 2), Ma Jinjun (1), Xu Dongjin (3), Chen Xuehao (1*), Jia Zhiming (3), Wang Yongli (3) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1257 Brassica campestris L. ssp. Chinensis var. utilis Tsen et Lee belongs to Cruciferae Brassica and is native to China, and is one of important vegetables mainly grown in the wide area of South China. Researches on male sterility gene of Brassica campestris L. ssp. Chinensis var. utilis with molecular makers play an important role in innovation of germplasmsa and breeding for new varieties. It has not been reported that ISSR is used to analysis male sterility gene of Brassica campestris L. ssp. Chinensis var. utilis. The genomic DNA of male sterile line and its maintainer line of Brassica campestris L£®ssp£®Chinensis var. utilis was studied with ISSR technique. The result showed that only primer UBC843 amplified different polymorphism band between sterile line and its maintainer line. The specific fragment was cloned and DNA sequence was determined. The size of specific fragment was 462 bp. Comparison between the specific fragment and other specific sequence derived from Chinese cabbage revealed that the homology generally was below 50%. According to the result sequenced, one pair of primers was designed to develop a more stable SCAR marker from the ISSR marker. The specific fragment amplified by ISSR primers was likely from nuclear DNA by F₂ population. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Shao Gaoneng, Tang Shaoqing, Luo Ju, Jiao Guiai, Tang Ao, Hu Peisong( *) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1252 A F7 recombinant inbred population consisting of 190 lines, derived from cross of a javonica (D50) and an indica (HB277) rice was used for QTL analysis for flag leaf morphological traits and grain shape in Hangzhou and Hainan. A total of 29 QTLs for flag leaf and grain shape were detected on 12 chromosomes except chromosome 2 and chromosome 11, and 6 QTLs were detected in two different growing environments and the contribution of these QTLs are more than 10%. Main-effect QTLs for flag leaf area¡¢flag leaf width and grain shape were detected on chromosome 1¡¢4 and 7, respectively. At the same time, these regions were also with QTLs for other traits, and they acted as QTL clusters, so three residual heterozygous lines of above region were selected for candidate region as gene genetic dissection¡¢cloning and molecular design breeding. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Hu Fengrong, Liu Guangxin, Xi Mengli, Wu Zhuhua, Shi Jisen( *) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1260 The chromosomes from root tips of Lilium sulphureum Baker were analysised by Giemsa C-banding. The result shown that, the karyotype is 2n=2¡Á=24, and the different C-banding points were shown on each chromosome. The C-banding karyotype would be: 2n=24=2CI+2I+4CI₊+2CI⁺+8I⁺+2I₊T₊+2IT₊+2I₊N. Two different depth centromeric bands and one secondary constriction band were shown on chromosome F. Each chromosome of Lilium sulphureum Baker can be distinguished by Gemisa C-banding analysis. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Zhu Yantao (1,2*), Li Dianrong (2), Guo Aiguang( 1), Tian Jianhua( 2) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1251 PolimaAand Shan 2A are two of the classic and effective cytoplasmicmale sterile (CMS) lines in Brassica napus L., and these two famous CMS lines have played very important roles in study and utilization of rapeseed heterosis in China even in the world. In early years, studies on the two CMS lines were mainly focus on the aspects of morphology and anatomy, cytology, physiology and biochemistry, genetics as well as utilization of heterosis. Where as in recent years, studies on them were mainly concentrated on the respect of molecular biology. This paper summarized the study states of Polima A and Shan 2A at home and abroad from above aspects, and introduced the new research progress on comparison of the two CMS lines in rapeseed in these years and their applications, existent problems and solving approaches in production and practice in rapeseed. With development of techniques of molecular biology, the research progress on the CMS lines in rapeseed will be more abroad and thorough. The further studies on PolimaAand Shan2Awill have an important significance to probe into themolecular mechanism of CMS and use the two CMS lines to service for scientific research and production in rapeseed. Thu, 28 Jan 2009 13:40:45 GMT Invited Review Zeng Jianmin (1), Yao Heng (1), Li Tianfu (1*), OuyangWentao (2), Cao Zhongxiang (1) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1261 Real time supervision of leaf chlorophyll content is necessary for better understanding the status of nitrogen nutrition and leaf maturity in flue-cured tobacco. This experiment was conducted in 2008 crop season at Yanhe station of Yunnan Tobacco Research Institute in Yuxi. In order to seek the best agent for plant chlorophyll extraction and determinate the relationship between chlorophyll content and SPAD readings, two varieties, K326 and Honghuadajinyuan (HD), were used in this study. Significant differences were observed in four agents for chlorophyll extraction in K326 and HD. The agent A, mixed acetone and ethanol with the volume ratio of 2 to 1 and then diluted to 95%solution, in which the chlorophyll content was 2 folds more than that in 80%acetone after 2 h extraction, extracted more chlorophyll than other three and showed higher extraction ability, rapider speed and greater efficiency in K326 variety. Moreover, the agent A, keep chlorophyll more stability than other three agents. The chlorophyll content, extracted by the agent A, was increased in extraction for 4 days than that for 1 day and decreased after 7 days in K326. However, it was lower in extraction both for 4 days and 7 days than that for 1 day in other three agents, especially for 95% ethanol and 80% acetone with the decreases of 1.24%~4.82%. Above similar findings were also observed in HD variety. Chlorophyll content in leaves was positively correlated with SPAD readings 50 days after transplanting (DAT) (R²=0.799 4^**) and 67 DAT (R²=0.866 2^**) in K326 and 68 DAT (R²=0.883 4^**) in HD. The whole leaf SPAD reading was the mean of 6 readings, of which each three data were measured at the two sides of the midrib in the middle of the full expansion leaf. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Zhong Huaiqin( 1,3), Wu Jianshe (1,3), Chen Shilin (2,3*), Chen Yuanquan (1,3), Lin Bing (1,3), Ye Xiuxian (1,3), HuangMinling (1,3) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1263 In this study, 16 primers were screened from 90 ten-bp random ones to carry out the polymorphic investigation of DNA-leveled on the germplasm resources of 16 ornamental sunflower by RAPD marker technique, and a total of 242 DNA fragments were amplified ranging from 200 bp to 2 800 bp, among which 162 ones (66.9%) were polymorphic, indicating abundant genetic diversity among the above ornamental sunflower idioplasms. The obtained results of genetic similarity analysis of the 16 ornamental sunflower genotypes showed that the Nei's coefficients ranged from 0.36 to 0.84, averaging at 0.590 3. Moreover, the cluster analysis distinguished all the tested genotypes to some extent and showed that the genetic classifications based on RAPD markers of most germplasm resources were relatively consistent with their origins. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Chang Cheng (1,2), Zhang Haiping (1,2), Zhang Xiuying (1), Yan Changsheng (1), Xiao Shihe( 1*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1262 PEBP-like gene is associated with grain size and weight in common wheat. In our study, the gene was assembled with two wheat ESTs and three barley ESTs screened based on the GS3 sequence of rice in GenBank. The relationship between allelic variation of PEBP-like and wheat kernel size and weight was researched through PCR using a pair of primers designed as the assemble sequence. The results were indicated that good correlation was found between grain length, width, thickness and weight. Grain width behaved the best positive correlation with thousand grain weight (0.668^**), the next was grain thickness (0.629^**) and grain length (0.485^**). Lines of RILs holding A type allele of PEBP-like showed smaller kernel and lower weight, on the contrary, lines with B type allele mean larger size and higher weight, which most of the lines were distributed the range of higher weight. Our results confirmed a close relation occurred in variation of PEBP-like gene and kernel size and weight of common wheat. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Hu Tingting (*), Liu Chao, Wang Jiankang, Ding Chengwei, Guo Rongliang, Wu Yuling, Xu Jiaan, Wang Youshuang http://www.molplantbreed.org/publish/abstract.asp?PaperID=1267 Rice is a kind ofmoderate-sensitiveness crop to salt, salt-tolerance characters belong to quantity character was controlled by polygene, and it is easy to be affected by environment etc. At present, salt tolerance QTLs which located in first, second, sixth, seventh chromosome mostlywere correlative to seedling salt tolerance; Salt tolerance varieties were cultivated by systemic selection, cross selection, stress tissue culture and identification; and the research future prospects in the futurewas discussed Thu, 28 Jan 2009 13:40:45 GMT Reviews Zhang Tao (1,3*), Zheng Jiakui (1,3), Wu Xianjun (2), Jang Kaifeng (1,3), Yang Qianhua( 1), ChenWenfu( 4), Yang Li (1,3) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1264 Protein content is important traits in evaluation of nutrient quality of rice. Most of crop traits are quantitative in nature, which are controlled by polygene. It's very important for rice nutrient quality breeding improvement to map QTLs of brown rice protein content. The objective of this study was to identify QTLs conditioning brown rice protein content using a recombinant inbred line (RIL) population derived from a cross between Zhongyouzao and Fengjin and its genetic linkage map by the composite interval mapping (CIM) of Windows QTL Cartographer 2.0 six QTLs (qPc-3¡¢qPc-6¡¢qPc-7¡¢qPc-8-1¡¢qPc-8-2 and qPc-11) showing significant additive effects for brown rice protein content were detected respectively. The six QTLs for brown rice protein content collectively explained 61.07% of the phenotypic variance, with the variance explained by a single QTL ranged from 3.79% to 19.41%. QTLs located in the vicinity of marker OSR34 on chromosome 8 displayed a major effect for protein content. Utilization of the QTL mapping on marker assisted selection for the improvement of grain quality in rice was also discussed. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Zhang Mingke, Zhang Lugang (*), Gong Zhenhui, Hui Maixia http://www.molplantbreed.org/publish/abstract.asp?PaperID=1265 Agenetic linkagemap of Chinese cabbagewas constructed, whose aimwas that the quantitative trait locis (QTLs) of purple trait etc were located. The map was constructed with JoinMap 3.0 software based on 231 polymorphicmarkers, using DNA samples extracted from a F2 population of purple Cai-tai (B. compestris L. var. purpurea Bailey) ¡ÁChinese cabbage (B. campestris L. ssp. Pekinensis (Lour.) Olsson), which composed of 11 linkage groups and 4 segments with 163 markers, included 117 RSAPs, 38 SRAPs, 5 SSRs and 3 RAPDs, covered genome 821.3 cM. The average distance between markers was 5.04 cM. A totle of 23.31% distorted markers distributed in the map and congregated in the linkage groups. Based the RAPD markers linked to purple trait, a conclusion that the 4th linkage group (LG4) corresponding to the 1st chromosome of Chinese cabbage was deduced. The map would be used effectively to QTL location of purple trait etc. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Peng Aihong( 1,3), Peng Zhuchun (1,3), He Yongrui (1,2,3), Xu Lanzhen (1,3), Lei Tiangang (1,3), Liu Xiaofeng (1,3), Yao Lixiao (1,2,3), Chen Shanchun (1,2,3*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1258 With the mature axillary buds of citrus excellent variety Shinanui (Citrus Reticulate¡ÁCitrus sinensis) severed as the explant, The plantlets which carried the hepatitis a virus capsid protein fusion gene were obtained through Agrobacterium-mediated transformation. In this study, survival in graft of mature axillary buds of shinanui was the highest, up to 90%, when the orange rootstocks were cultured 15 days. Sterilization time was effected on maturity of shinanui branches. The more maturity of branches, the longer of sterilization time was. The putative transgenic plantlets was selected by PCR, PCR amplification product of PCR positive plantlets were sequenced, and the results showed that Hepatitis a virus capsid protein fusion gene was integrated into genome of transgenic shinanui. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Jiang Chunya, Liao Jiao, Xu Xiaobiao (*), Gu Qingqing, Liu Shanjun, Chen Jinyin http://www.molplantbreed.org/publish/abstract.asp?PaperID=1273 Expressed sequence tags (ESTs) are short, usually unedited sequences obtained by single-pass sequencing of cDNA clones from any cDNA library. Currently, plant EST collections comprise over 15 million sequences from about 200 species. A number of studies indicate that EST analysis is one of the most effective approaches to explore big genomes. Analyzing and comparing ESTs can provide in formation on gene expression, function and evolution. Large-scale EST sequence has become an attractive alternative to discovery plant genome sequence and plant metabolism analysis. The research status based on the plant EST-SSR markers was introduced, and the genetic map construction, genetic diversity and generality by using the EST-SSR technology in some crops were also reviewed. At the same time, EST also made use of a number of deficiencies and development trends. Thu, 28 Jan 2009 13:40:45 GMT Reviews Zhang Yadong, Hu Xingyi (*), Song Congwen http://www.molplantbreed.org/publish/abstract.asp?PaperID=1268 The increasing availability of expressed sequence tags (EST) in public databases provides a valuable resource for development of new EST-SSR molecular markers. In this study, the 8 Populus varieties from Hubei province were identified by EST-SSR markers, and the genetic parameters on different loci were analyzed. The result about variety identification showed that only 4 pairs of EST-SSR primers could totally differentiate 8 Populus varieties, and the genetic diversity analysis showed that polymorphic loci percentage was 58.3%, average effective number of alleles was 2.09, average Shannon's information index was 0.61, and average expected heterozygosity was 0.35. All these results indicated that EST-SSR markers from Populus can be used for the analysis of variety identification and population genetic diversity. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Zhang Miaobin, Pan Lijing (*), Fan Ganqun, Chen Jimin, Cheng Ping( *) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1286 Isolation of high-quality genomic DNA of transgenic Dendrobium plantlets for rapid screen or genetic analysis by PCR amplification and Southern blot is difficult, for the transgenic plantlets grow very slowly and contain high-level polysaccharides. In order to overcome such problems as low yield, degradation, poor PCR amplification and poor restriction digestion, three methods developed from hexadecyltrimethylammonium bromide (CTAB) or sodiumdodecyl sulfate (SDS)method respectivelywere optimized and compared in this paper. Of the three modified methods used, method¢ò produced pure and high-quantity genomic DNA from small amount samples of transgenic Dendrobium or other plantlets in vitro tissue culture. DNA isolated by method¢òwas proved amenable to PCR amplification, restriction digestion and Southern blot analysis of transgenic Dendrobium plantlets. Thu, 28 Jan 2009 13:40:45 GMT Novel Thinking & Technologies Zhang Haiping, Feng Jiming, Yin Bo, Chang Cheng (*), Si Hongqi Ma Chuanxi http://www.molplantbreed.org/publish/abstract.asp?PaperID=1259 Pre-harvest sprouting in wheat significantly reduces end-use quality, seed value and yield. In this study, recombinant inbred lines (RILs) populations (Wanxian Baimaizi/Jing411) were used to detect validation ofmolecular marker mapped on the 3rd (XBarc321, XBarc310 and XBarc57) and 4th chromosomes (Xbarc170, Xgwm397 and Xgwm269) closely linked to pre-harvest sprouting resistance gene reported in previous study. The results indicated that molecular marker (XBarc321, XBarc310 and XBarc57) located on the 3rd chromosomes were more effective than molecular marker located on the 4th chromosomes. These results provide important information for mapping QTL loci associated with pre-harvest sprouting resistance in Wanxian Baimaizi/Jing411 populations. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Li Lubin (1), Liu Lei (2), Yuan Jinling (3), Dong Yinmao (2), Hu Tao (1), Miao Kun( 1), He Congfen( 2*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1269 Themethod of CTAB was used to extract 37 species bamboo genomic DNA. The primers were designed based on the chloroplast 5S rDNA internal transcribed spacer (ITS) and matK gene sequence which were published in the GeneBank. The PCR product of 5S rRNA ITS is about 450 bp and then was sequenced directly. The sequencing results were analysed by software DNAMAN and showed that the 37 species had the same length except Phyllostachys shuchengensis had a variable site (compared with other species among Phyllostachys) in the 392nd (A¡úC). The results indicate that 5S rDNA ITS sequence in 37 species bamboo was very conservative and it could not provide enough information for the relationship of the species among Phyllostachys. PCR product of matK sequence amplified from 3 species bamboo genomic DNA was sequenced respectively and it is about 1 500 bp. Based on multiple sequence alignment of matK sequence, RFLP (restriction fragment length polymorphism) site (BstN¢ñ) was discovered. After all the PCR products of matK sequence were digested by BstN¢ñ, three species Phyllostachys meyeri, Phyllostachys parvifolia and Phyllostachys angusta were branched from the others in Phyllostachys in the 938th. The results indicated that the matK gene in Phyllostachys is also conservative, and it cannot provide enough information for evolutionary studies under Phyllostachys. The chloroplast 5S rDNA ITS gene band matK gene have no value for the phylogenetic relationships in the genus of Phyllostachys, but they could be used to study the phylogenetic relationships of above the genera and faster rate of evolution in the genera. Thu, 28 Jan 2009 13:40:45 GMT Research Reports Zhou Kanshe (1), Liu Yilan (1), Wang Yanrong (2*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1270 Genetic diversity of seed yield char acteristics was studied with different varieties of alfalfas using morphological, a field study was conducted from 2005 to 2006 in Linze county of Zhangye city, Gansu province, genetic diversity of seed yield and its components were investigated in 62 alfalfa germplasm resources belonging to 55 varieties which grew in the fourth year under row sowing condition. Variation of seed yields was large among varieties, and the two years results also showed that each of the seed yield components among varieties and between years vaired greatly. All the seed yield components varied among varieties, and there were significantly differences between domestic varieties and introduced varieties except number of seeds per pod. The number of inflorescences per fertile shoot was significantly correlated with seed yield in two years, it was stressed that the number of inflorescences fer fertile shoot could be an useful character to be used as a selection criterion for increasing the seed yield potential of alfalfa row sowing condition. The mean percentage of actual seed yield accounted for potential seed yield (ASY/PSY) was less than 4% in two years, most of them was between 1% and 2%, and difference significantly was found among varieties and between years. It was found that 1 000 seed weight was different significantly among all domestic sources, but not different in introduced sources (p>0.05). Thu, 28 Jan 2009 13:40:45 GMT Research Reports Niu Yanbing (1*), Qing Ling( 2*), Wang Defu (1), YaoMin (1), Guo Pingyi( 1**) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1271 RNA silencing is a sequence-specific RNA degradation system that is conserved in eukaryotic cell. In high plants, RNA silencing serves as an adaptive antiviral defense system that is activated by, and targeted against, viruses. As a response to the defense system, viruses have evolved suppressor proteins that act at different points in the silencing pathway and ultimately overcome the host silencing response. In this paper, we discuss the current methods available for identification and preliminary analysis of silencing suppressors and their putative modes of action. We also discuss the biological value of silencing suppressors in plants including the probing RNA silencing pathway, the increase expression levels of protein and the developmental anomalies induced by silencing suppressors. Thu, 28 Jan 2009 13:40:45 GMT Reviews Wang Jiani, Dai Xiaofeng( *) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1272 Advances in mechanism and risk of the resistance for Helicoverpa armigera to transgenic Bt cotton was reviewed, as one important reason for the Bt cotton resistance management. The management strategies and techniques of resistance for Helicoverpa armigera to the transgenic Bt cotton, included gene strategies (such as insert a set of insecticidal gene, tagged and organ-specific expression, infection inducing expression, regulating expression, high dose and low dose expression strategy), field tactics (such as the rotation and mixture planting of the Bt cotton, reduced the economic threshold for chemical control, cultivation measures and the refuge tactics), and the state macro-adjustment (such as implementation of the zoning planting system, strengthening management of seed production, viewing the resistant dynamic monitored, and giving strict safety evaluation for Bt cotton planting) were discussed mainly. Thu, 28 Jan 2009 13:40:45 GMT Reviews Zhang Zhengfeng (1), Xiao Benze (2*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1274 The genome sequencing and expressed sequenced tag (EST) projects generated a vast amount of sequences data. Simultaneously, the increasing bioinformatics software facilitates the development ofmolecular markers. Additionally, to develop molecular markers in laboratory will benefit from the advanced biotechnology and construction of batch of operation systems. The methods, characteristics and corresponding application of developing SNP and SSR based on bioinformatics and developing SSR, STAP, TRAP on biotechnology were reviewed. The progress of conversion of RAPD and AFLP to single-locus specific PCR markers were also presented. Finally, how to select suitable technique for different plants and develop functionalmarkers are elucidated. Thu, 28 Jan 2009 13:40:45 GMT Reviews Chen Yajuan, Li Fuguang (*), Liu Chuanliang, Zhang Chaojun, Wu Zhixia, Zhang Xueyan http://www.molplantbreed.org/publish/abstract.asp?PaperID=1275 Osmotic adjustment is one of the most important ways that higher plant devolepes to maintain the osmotic balance and resist to environmental stress under water deficiency. Over-expression of certain genes which encode key enzymes involved in the osmolytes biosynthesis, showed improved drought tolerance in transgenic plants£¬which suggests a potential application of these genes in cotton genetic improvement. This article reviews the advance in several important osmoprotectants based on the current research and discusses the promising application of osmoprotectant in enhancing cotton drought tolerance and molecular genetic improvement. Thu, 28 Jan 2009 13:40:45 GMT Reviews Liao Yi( 1,2), Sun Baojuan (2), Sun Guangwen( 1), Liu Houcheng( 1), Chen Riyuan( 1*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1276 Identification of molecular markers linked with gene can promote the development of MAS (molecular assistant selection) and map-based cloning. Bulked segregant analysis (BSA) as a classical method for rapidly and effectively identifying markers linked to any specific gene or genomic region was widely used in crop breeding study. BSA also has several advantages over the use of near-isogenic lines (NILs). In this article the basic principle and theory of BSA were introduced, and several key problems about using BSA were analyzed, including the effects of genome size of different species, markers unlinked to the aimed loci appeared differences between bulks and construction of the ideal DNA pools, etc. At the same time the applications of the method in the different segregating generations such as the F₂ ¡¢backcross¡¢RIL¡¢DH and so on were introduced and compared. Finally we discribed the application prospects of BSA to be used for quantitative analysis between DNA pools. In conclusion we wish to offer some academic references for the application of BSA by our introduction and analysis. Thu, 28 Jan 2009 13:40:45 GMT Reviews Zhang Hechen( 1), Ye Chuyu (2), Xia Xinli (2*), YinWeilun (2*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1277 In plant, intracellular calcium levels altered in response to multiple abiotic stresses result in calcium signatures. Although the specific signature of the calcium transients can encode information on its own, an additional level of regulation in calcium signaling is achieved via the action of calcium-binding proteins. Calcineurin B-like proteins, as the plant calcium-binding proteins recently identified, must function by binding and regulating a group of serine-threonine protein kinases called CIPKs (CBL-interacting protein kinases) and provide a signal network to adapt various stresses. The calcium signal pathways have been well studied in plant and it has been identified that the pathways play important roles in multiple abiotic stresses, including salt, drought and low K⁺. Thu, 28 Jan 2009 13:40:45 GMT Reviews Yao Danqing, Zhang Weiwei, Yuan Lihua, Pan Junsong, He Huanle, Cai Run (*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1278 Virus-induced gene silencing (VIGS) is a technology that exploits an RNA-mediated antiviral defense mechanism and has been shown to be of great potential in plant reverse genetics. Because of the methodological simplicity, robustness, and speedy results than plant normal transformation, VIGS has been used widely for analysis of gene function and high-throughput functional genomics in plant. Until now most applications of VIGS have been in Nicotiana benthamiana. Moreover, new vector systems and methods are developed to use in other plants, including monocots. Here, we discuss the issues specific to the application of VIGS technology to determine gene function, which has revealed the roles of a variety of genes in disease resistance, abiotic stress, cellular signaling and secondary metabolite biosynthesis and then construct a series of new related models. Thu, 28 Jan 2009 13:40:45 GMT Reviews Xie Chunzheng (1,2), LiWanfu (1,2), Liu Haiyan (1), Li Ling (2), Liang Xuanqiang (1*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1280 To characterize the function of the AhPIP1 (pathogenesis-induced protein gene from Arachis hypogaea L.), we have isolated AhPIP1 (EU860093) gene by RACE and promoter region by GenomeWalking method from peanut and have been expressed the AhPIP1 in BL21 in this paper. As result, the AhPIP1 gene encodes a 193-amino-acid polypeptide, exhibiting high similarity with other members of PIP, and the open-reading flame is 579 bp. Genome amplification sequence is 1 883 bp, including three exons and two introns, length of 602 bp and 600 bp respectively. The AhPIP1 promoter sequence about 513 bp which were received by Genome Walking method contain DRE¡¢MYB/MYC¡¢GARE¡¢SEBF core sequences or other abiotic-responsive cis-acting elements and the result indicate the AhPIP1 gene may play important role in peanut stress responses and growth and development. we obtained the recombination AhPIP1 protein, expressed in BL21 about 40 kD in SDS-PAGE gel. Cloning the gene and expressing the fusion protein has laid the foundation for the function study of AhPIP1. Thu, 28 Jan 2009 13:40:45 GMT New Gene & Germplasms Guan Ning, Li Cong (*), Wang Yongxin, Miao Lihong http://www.molplantbreed.org/publish/abstract.asp?PaperID=1279 The cDNA of Na⁺/H⁺ antiporter gene from Elytrigia trichophora named EtNHX1 (Accession numeber: EU876834) was isolated by RT-PCR, with primers designed according to the sequence of TaNHX1 and TeNHX1. The largest open reading frame of EtNHX1 gene has 1 641 bp in length and encoded a polypeptide of 546 amino acid residues. The estimated molecular weight and isoelectric points of the putative protein were 59.8 kD and 8.0, respectively. Components of amino acids encoded by EtNHX1 contained 39 basic amino acids, 37 acidic amino acid, 256 hydrophobic amino acids and 128 polar amino acids. The predicted secondary structure composition for the protein has about 47% alpha helixes, 20% extended strand, 4.5% beta turn and 28% random coil. Hydrophobicity/ hydrophilic analysis indicated that the EtNHX1 had 12 hydrophobic segments containing 10 potential transmembrane segments. Sequencing analysis showed that EtNHX1 had high identity with TaNHX1, TiNHX1, TeNHX1, OsNHX1, ScNHX1, ThNHX1, and had about 98%, 98%, 96%, 85%, 68%, and 67%similarity respectively. Blast result and the phylogenetic tree analysis showed that EtNHX1 was Na⁺/H⁺ antiporter located on vacuolar of Elytrigia trichophora. Thu, 28 Jan 2009 13:40:45 GMT New Gene & Germplasms Cheng Yu (1), Lin Ruiming (2*), Ouyang Hongyu (3), Lv Shuxia( 1), Xu Shichang (2) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1281 Wheat yellow rust, caused by Puccinia striiformis f. sp. tritici, is one of the plant foliar diseases spreaded through air around the world, and it often happens severely in China and causes great yield loss on wheat (Triticum aestivum L.). It is very important to clone and characterize biological functions of the resistance-relatedwheat genes to yellow rust in practical utilization. In this study, we constructed a wheat cDNA library using the near-isogenetic line Taichung 29*6/Yr5 seedlings challenged by incompatible yellow rust race CY32. The results showed that the primary library titer is 0.9¡Á106 pfu/mL, and 1.0¡Á109 pfu/mL for amplified library, and 98% recombinants were obtained. The inserted cDNA fragment size of this library is between 0.5 kb to 3 kb, and most cDNA clone is about 1.0 kb in length. Therefore this cDNAlibrary is qualified forwheat gene cloning and function analysis. Thu, 28 Jan 2009 13:40:45 GMT New Gene & Germplasms Liang Xiaolian (1,2), Li Huiliang (1), Peng Shiqing (1*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1282 By using the information about the sequence from a differentially expressed clone (designated as Hb-SSH12) encodes a translationally controlled tumor protein (TCTP) that was cloned recently by the subtractive hybridization between in latex of Hevea brasiliensis juvenile clone and in those of Hevea brasiliensis mature clone, a full-length cDNA encoding TCTP, designated as HbTCTP, was isolated and characterized from rubber tree. Sequence analysis revealed that the ORFs of HbTCTP encode 168 amino acid residues with a total predicted molecular mass of 19.14 kD. The deduced amino acid sequences of HbTCTP showed high identities of 93.45%¡¢90.48%¡¢89.29%¡¢85.12%¡¢82.74% and 79.76% to those of the TCTP from Jatropha curcas, Elaeis guineensis, Arachis hypogaea, Cucurbita maxima, Solanum lycopersicum and Arabidopsis thealiana. The result of RT-PCR analysis indicated that the transcripts of the HbTCTP were constitutively expressed in the latex, in the leaves and in the barks. The transcription of HbTCTP was induced by ethylene, suggesting that HbTCTP may involve ethylene signaling. The present study contributes towards an understanding of the molecular characterization of TCTP in rubber tree. Thu, 28 Jan 2009 13:40:45 GMT New Gene & Germplasms Lin Haimei( 1,2), Guo Anping (2*), Wang Xiaoling( 3), Guo Yunling (2), Kong Hua( 2), He Lika (2) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1285 To explore the extraction of total RNA from Oryza longisatminata, the author attempt modified SDS method, CTAB method and TRIzol method, comparing the extraction effect of these methods, by electrophoresis in general agarose gel and spectrophotometer readings. The results indicates that modified SDS method, CTAB method for root RNA is good, its bonds of electrophores are clear, there are almost no contamination, no obvious degradation, the OD₂₆₀/OD₂₈₀ is between 1.75~2.0, it can meet the general equirements of molecular operation, but the products of RNA from leaf are polluted by more DNA, and a little proteins total RNA with the TRIzol method are polluted by proteins, with different extent, no matter root or leaf such RNA must be purified if it will go to the next molecule operation. That is to say, modified SDS method and CTAB method is the first choice for root total RNA. Thu, 28 Jan 2009 13:40:45 GMT Novel Thinking & Technologies Li Yanjun, Zhang Xinyu, Yang Huina, Tian Xinhui, Sun Jie (*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1283 A subtracted cDNA library was constructed via suppressive subtractive hybridization (SSH), in which the brown colored cotton Xincaimian 5 was utilized as tester while driver is its white colored susceptible near-isogenic line NIL-C5. Total 400 clones were identified from screening of the subtracted library by PCR, and further verified by Reverse Northern Dot-Blot. 20 differentially expressed clones were selected for sequencing analyses, of which the results were aligned with GenBank databases via BLASTX programs. It is shown that 5 of 20 candidates were identity with known genes in GenBank, 1 of them showed significant similarity to 4CL, a important gene for pigment synthesis, and no similar sequence were found in GenBank for the other fragments. Thu, 28 Jan 2009 13:40:45 GMT New Gene & Germplasms Wang Yanqing, Ji Kongshu (*) http://www.molplantbreed.org/publish/abstract.asp?PaperID=1284 In this paper, the orthogonal design was used to optimize SRAP-PCR amplification system on Paeonia suffruticosa on four levels of five factors (Taq DNA polymerase, Mg²⁺, DNA template, dNTPs, and primer, respectively), The results showed the order of each factor in different levels affected on the result of PCR was Taq DNA polymerase, primer, dNTPs, Mg²⁺ and DNA template. A most suitable SRAP-PCR system for Paeonia suffruticosa was established that was total 25 ¦ÌL reaction system containing 0.5 U Taq DNA polymerase, 2.0 mmol/L Mg²⁺, 50.00 ng DNA template, 0.20 mmol/L dNTPs, and 0.30 ¦Ìmol/L primer. This optimized system for SRAP marker would become one of the protocols for further research on Paeonia suffruticosa. Thu, 28 Jan 2009 13:40:45 GMT Novel Thinking & Technologies