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10 November 2010, Volume 24 Issue 6 Previous Issue    Next Issue
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研究报告
综述与专论
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实验技术


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研究报告

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Analysis on SterilityRelated Genes in a New Type of Japonica Photo(thermo)Sensitive Male Sterile Line under Short Daylength and Low Temperature Conditions by Microarray DU Shi-yun,YANG Jing,WANG Shou-hai,WANG De-zheng,WU Shuang,LUO Yan-chang,LI Yang-sheng 2010, 24(6): 559-566 .  DOI: 10.3969/j.issn.1001-7216.2010.06.001 Abstract ( )   PDF (2960KB) ( )   A new type of japonica photo(thermo)sensitive cytoplasmic male sterile line 2310SA was bred. This line contains photo（thermo）sensitive genic male sterile（PTGMS） genes in the twoline system hybrid rice and cytoplasmic male sterile (CMS) genes in the threeline system hybrid rice, which has a great application potential in hybrid rice breeding for its stable sterility. To understand the underlying genetic mechanism and to explore male sterility of rice, the selected 2310SA line and its maintainer line of the twoline system, PTGMS line 2310S, were grown in Hefei, China. The anthers were collected at four panicle developmental stages (meiosis, mononucleate, binucleate and mature stages) and the isolated RNA were used for microarray analysis. Results showed that differentially expressed genes(DEGs) at meiosis stage(1938 genes upregulated, 1635 genes downregulated) and mature stage(2220 genes upregulated，2656 genes downregulated) were more than those at mononucleate stage(752 genes upregulated, 693 genes downregulated) and binucleate stage(1025 genes upregulated, 886 genes downregulated). The number of downregulated genes was higher than that of upregulated genes at mature stage, whereas at other stages the number of upregulated genes was more than that of downregulated genes. Moreover, hierarchical cluster analysis of all DEGs suggested that the expression of the DEGs at mononucleate and binucleate stages belonged to the same cluster, and the expression of the DEGs at meiosis and mature stages to another two clusters. Among DEGs, 147 genes were found to be differentially expressed at all the stages when comparing 2310SA line with the control line 2310S. For the 147 common DEGs, the downregulated genes were more than the upregulated ones at mature stage but not at other three stages. Additionally, these common DEGs were involved in many biological processes, which were classified into 11 functional categories, including biosynthesis, response to stimulus, photosynthesis, macromolecule metabolic process, signal transduction, regulation of transcription, carbohydrate metabolic process, flower development and cell death. Among proteins coded by the common DEGs, cytochrome P450 family protein and betaring hydroxylase play important roles in fatty acid metabolism, and heat shock transcription factor 29, heat shock protein DnaJ family protein and heat shock protein 70 are involved in stress response. These genes are likely to relate directly or indirectly to cell death and pollen abortion.

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Molecular Analysis of Rice CIPKs Involved in Biotic and Abiotic Stress Responses CHEN Xi-feng,GU Zhi-min,LIU Feng,MA Bo-jun,ZHANG Hong-sheng 2010, 24(6): 567-574 .  DOI: 10.3969/j.issn.1001-7216.2010.06.002 Abstract ( )   PDF (3888KB) ( )   Plant calcineurin Blike (CBL) proteins have been proposed as important Ca2+ sensors and specifically interact with CBLinteracting protein kinases (CIPKs) in plantspecific calcium signaling. We identified and isolated 15 CIPK genes in japonica rice variety Nipponbare based on the predicted sequences of rice CIPK gene family. Gene structure analysis showed that these 15 genes were divided into intronless and intronrich groups, in which OsCIPK3 and OsCIPK24 exhibit alternative splicing. The phylogenetic analysis indicated that rice CIPKs share an ancestor with Arabidopsis and poplar CIPKs. Analysis of gene expression showed that these OsCIPK genes were differentially induced by biotic stress (bacterial blight, BB) and abiotic stresses, including heavy metal (Hg2+), high salt, cold and ABA. Interestingly, five OsCIPK genes (OsCIPK1, OsCIPK2, OsCIPK10, OsCIPK11 and OsCIPK12) were transcriptionally upregulated after BB infection, among which four OsCIPK genes (OsCIPK2, OsCIPK10, OsCIPK11 and OsCIPK14) were induced by all treatments, indicating that some of OsCIPKs were involved in multiple stress response pathways in plants. The findings suggest that CIPKs play a key role in both biotic and abiotic stress responses.

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Molecular Cloning and Quantitative Expression of a FullLength cDNA Encoding β1Tubulin in the Ovary of Rice Water Weevil, Lissorhoptrus oryzophilus (Coleoptera: Curculionidae) YU Kui-feng,LI Hong-liang,YANG Pu,CUI Xu-hong,ZHU Zeng-rong,SHANG Han-wu 2010, 24(6): 575-580 .  DOI: 10.3969/j.issn.1001-7216.2010.06.003 Abstract ( )   PDF (4145KB) ( )   A fulllength cDNA encoding β1tubulin from the parthenogenetic rice water weevil (RWW), Lissorhoptrus oryzophilus in China and from the sexual RWW in the United States was cloned by RACE and RTPCR, respectively. The full cDNA sequence was 1727 bp for the parthenogenetic RWW, and 1730 bp for the sexual RWW. Both cDNA sequences contained a 1344 bp open reading frame encoding 447 amino acid residues. The putative molecular weight of these amino acid residues was approximately 50 kD at an isoelectric point of 4.54. The cloned β1tubulin amino acids shared a high similarity (95% to 100%) with the βtubulin homologues from other insects. Absolute quantification revealed that the β1tubulin was expressed at a significantly higher level in the ovary of the parthenogenetic RWW (3.14×107 copies/μL±0.66×107 copies/μL) than the sexual RWW (5.08×106copies/μL±0.47×106 copies/μL). It is suggested that the differential expression of β1tubulin in the parthenogenetic RWW may play an important role in the formation of the mitotic spindle.

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Improvement of Gall Midge Resistance for Restorer Line in Rice by Molecular Marker Assisted Selection LIU Wu-ge ,WANG Feng ,XIAO Han-xiang ,LIU Zhen-rong,LI Jin-hua ,LIAO Yi-long ,ZHU Man-shan ,HUANG Hui-jun,HUANG Bing-chao ,ZHANG Yang 2010, 24(6): 581-586 .  DOI: 10.3969/j.issn.1001-7216.2010.06.004 Abstract ( )   PDF (442KB) ( )   Gm6, a dominant gene conferring resistance to the four biotypes of Asian rice gall midge (Orseolia oryzae WoodMansion) occurred in South China from the donor KG18, was introduced into the restorer line Guanghui 998 with good grain quality and poor gall midge resistance, by using backcross breeding and molecular markerassisted selection. Six elite improved lines were developed based on the results of molecular markerassisted analysis, the recovery ratio of genetic background and the performance of agronomic traits. Evaluation of resistance to Chinese gall midge biotype 1 and biotype 4 was carried out. All the improved lines were immune to the pest with the ratio of silver shoots of 0 whereas the check Guanghui 998 were highly susceptible with the ratio of silver shoots of 98%. The hybrid F1, derived from the improved lines, showed moderate resistance against gall midge with the ratios of silver shoots ranging from 10% to 20%. The investigation on agronomic traits of the improved lines showed that there was no significant difference between the improved lines and the check except for 1000grain weight between one line and the check Guanghui 998. The improved lines could be used for breeding hybrid rice with higher gall midge resistance.

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Creation and Characteristics of Indica Type Cytoplasmic Male Sterile Rice Line Gaoguang A Labeled with AlbinotoZebra Leaf Marker ZHANG Hong-lin,CHENG Jian-feng,LIU Yue-qing,ZHONG Xiao-ying,ZHANG Rui-xiang,LIU Hai-ping,ZHANG Pu 2010, 24(6): 587-594 .  DOI: 10.3969/j.issn.1001-7216.2010.06.005 Abstract ( )   PDF (3675KB) ( )   A new mutant with albinotozebra leaf color trait was selected from the progeny of the indica rice maintainer line D57B by irradiation with 300 Gy 60Co gamma rays. The mutant was then backcrossed with D57A as the recurrent parent, and finally a novel type of cytoplasmic male sterile rice line, Gaoguang A, labeled with albinotozebra leaf color marker was developed through continuous selection. The mutant trait was controlled by a single recessive nuclear gene, expressing albino (first to sixth leaves) and zebra leaves (from the sixth leaf) and greenish albino young panicles. Gaoguang A belongs to early maturity earlyseason indica rice, with a duration from seeding to heading of about 65 days. Compared with D57B, Gaoguang A had lower plant height and longer flag leaf, which are beneficial to pollination and fertilization, high seed setting rate and fully filled grain number of hybrids, whereas its seed germination rate was slower. The other agronomic traits of Gaoguang A were similar to D57B. Gaoguang A had some excellent out crossing characteristics, such as high stigma exsertion rate, strong vitality of stigma, long flowering time in a day, small tension angle of floret, low closed glume rate, sensitivity to gibberellins. It also had excellent combining ability with other varieties. In the mixed sowing experiment of Gaoguang A and hybrid rice combinations, the weaker competitiveness of young seedlings of Gaoguang A resulted in natural death of its young seedlings. Finally, the prospects of utilizing Gaoguang A in the threeline hybrid rice production were discussed.

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Genetic Improvement and Biological Characteristic Analysis of Dwarf Abortive Type Japonica Cytoplasmic Male Sterile Line with Wide Compatibility SONG Xin-wei,LIN Jian-rong,WU Ming-guo* 2010, 24(6): 595-600 .  DOI: 10.3969/j.issn.1001-7216.2010.06.006 Abstract ( )   PDF (1030KB) ( )   From the view of methodology for genetic improvement of japonica cytoplasmic male sterile (CMS) lines, the breeding of the dwarfabortive type(DA type) japonica CMS line Chunjiang 95A with wide compatibility was described. The biological characteristics of the DA type Chunjiang 95A and its isonuclear alloplasmic CMS line DianI type Chunjiang 95A were investigated. The results showed that the DA type Chunjiang 95A was a japonicalinous CMS line and mainly typical abortive, and had a better fertility stability than stained abortion type japonica CMS lines. The DA type Chunjiang 95A was featured by high stigma exsertion, good flowering habit, high outcrossing rate and wide compatibility. Its recoverability was different from stained abortion type japonica CMS lines. In addition, genetic improvement of the typical abortion type japonica CMS lines and the application prospects of heterosis were discussed.

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Estimation of Mating System in Natural Oryza rufipogon Populations by SSR Markers LI Xiao-xiang,LIU Yong,DUAN Yong-hong ,WANG Shu-hong ,ZHAN Qing-cai ,SUN Gui-hua ,GAO Li-zhi 2010, 24(6): 601-607 .  DOI: 10.3969/j.issn.1001-7216.2010.06.007 Abstract ( )   PDF (235KB) ( )   Oryza rufipogon Griff．is the most important germplasm for rice improvement．Better understanding its mating system will provide useful information for further in situ conservation and management of O. rufipogon． By using SSR method，the genetic diversity and outcrossing rate of six natural O.rufipogon populations were estimated. The levels of genetic diversity varied among the populations, with the Nei′s diversity indices ranged from 0.3166 to 0.6075. There was a significant genetic differentiation (Fst=0.422) among the sampled populations. Two populations exhibited a heterozygosity excess, whereas the other four populations had a heterozygosity deficit. The O.rufipogon populations possessed a mixed type of mating system, although the outcrossing rate was variable among the populations. The multilocus outcrossing rate ranged from 15.1% to 418%，and the singlelocus outcrossing rate from 10.1% to 28.5%．There were significant differences between the multilocus outcrossing rate and the singlelocus outcrossing rate, and high values of correlation of paternity in the populations, indicating a considerable inbreeding existed in the populations. The correlation of paternity for the multilocus significantly differed from the correlation of paternity for the singlelocus, suggesting that there was a substructure occurred in each sampled population. Some measures for further improving insitu and exsitu conservation of O. rufipogon were also discussed.

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Fitness of Progenies Between Transgenic Rice and Weedy Rice under Greenhouse Conditions ZUO Jiao,QIANG Sheng,SONG Xiao-ling* 2010, 24(6): 608-616 .  DOI: 10.3969/j.issn.1001-7216.2010.06.008 Abstract ( )   PDF (175KB) ( )   In greenhouse, the F1 and F2 hybrids derived from glufosinateresistant transgenic rice (Y0003 and 991) and Malaysian weedy rice or Anhui weedy rice were backcrossed with corresponding weedy rice to obtain backcross generations. And the seed setting rates of the backcross were counted. The proportions of plants carrying the glufosinateresistance gene in F1 or F2 hybrids and the backcross generation were calculated. In order to comprehensively assess the risk of gene flow from glufosinateresistant rice to weedy rice, the fitness of hybrid and backcross generations were observed. The results showed that the hybrid generations were able to backcross with corresponding weedy rice with seed setting rates from 15% to 60%. The seeds from reciprocal hybrid F1 generation showed favorable glufosinateresistance, but F2 and backcross generations showed a normal 3∶1 and 1∶1 Mendelian segregation with glufosinate selection. The fitness of hybrid and backcross generations didn′t differ obviously from that of their corresponding weedy rice. Most hybrid generations displayed similar fitness with backcross generations, but some hybrid generations were lower than backcross generations in fitness. All above results indicate that hybrids and backcross generations of two weedy rice carrying glufosinateresistance genes of Y0003 and 991 could survive under benign conditions without herbicide selection.

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Establishment of Agrobacterium tumefaciensMediated Transformation System for Rhizoctonia solani AG-1 ⅠA, the Causal Agent of Rice Sheath Blight YANG Ying-qing#,YANG Mei#,LI Ming-hai,LI Yong,HE Xiao-xia,ZHOU Er-xun* 2010, 24(6): 617-622 .  DOI: 10.3969/j.issn.1001-7216.2010.06.009 Abstract ( )   PDF (2118KB) ( )   In order to construct the TDNA insertional mutagenesis transformation system for rice sheath blight pathogen Rhizoctonia solani AG1 ⅠA, the virulent isolate GD118 of this pathogen was selected as the initial isolate for transformation. The conditions for the transformation of GD118 were optimized in 5 aspects, i.e. preinduction time, coculture time, acetosyringone concentration at coculture stage, coculture temperature and pH values of solid induction medium (SIM) at coculture phase. Finally, an Agrobacterium tumefaciensmediated transformation (ATMT) system for R. solani AG1 ⅠA was established successfully. The optimal conditions for this ATMT system are as follows: the concentration of hygromycin B at 30 μg/mL for transformant screening, 8 h of preinduction time, 20 h of coculture time, 200 μmol/L of acetosyringone in SIM at coculture stage, coculture temperature at 25℃ and pH 5.6 to 5.8 of SIM at coculture phase. The transformants still showed high resistance to hygromycin B after 5 generations’ subcultures. Ten transformants were randomly picked out for PCR verification using the specific primers designed from the hph gene, and the results revealed that an expected band of 500 bp was amplified from all of the 10 transformants. Meantime, PCR amplification for these 10 transformants was carried out using specific primers designed from the Vir gene of A. tumefaciens, with 4 strains of A. tumefaciens as positive controls for eliminating the falsepositive caused by A. tumefaciens′s contamination, and the results showed that an expected band of 730 bp could be amplified from the 4 strains of A. tumefaciens, whereas no corresponding DNA band could be amplified from the 10 transformants. The results of above two PCR amplifications clearly showed that TDNA was indeed inserted into the targeted isolate GD118.

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Pathogenicity Differentiation of Rice Sheath Blight Pathogen Rhizoctonia solani AG-1 ⅠA Isolates from Anhui and Hubei Provinces, China WANG Ling,HUANG Wen-wen,HUANG Shi-wen,LIU Lian-meng,LIU En-yong, 2010, 24(6): 623-629 .  DOI: 10.3969/j.issn.1001-7216.2010.06.010 Abstract ( )   PDF (1273KB) ( )   Pathogenicity variation of 200 isolates of rice sheath blight (SB) pathogen Rhizoctonia solani AG1 ⅠA from the infected samples from provinces of Anhui and Hubei, China were analyzed by using five cultivars with different resistance to SB at the seedling stage in greenhouse. There were significant differences in pathogenicity among these tested isolates of R. solani AG1 ⅠA. The frequency of average disease index followed a normal school distribution. Based on their disease indices to the five cultivars, the isolates were classified into three distinct pathotype groups by dynamic cluster analysis, i.e. weak, moderate and strong pathotypes, accounting for 29.5%, 60.5% and 10.0%, respectively. The moderate pathotypes were dominant. Random distribution of pathotypes in different areas implied that R. solani AG1 ⅠA population was a mixture of pathotypes naturally. The Mantel testing revealed that the pathogenicity variation was not obviously correlated with the collective location. The discriminant functions of the three pathotypes were constructed based on the Bayes method. The accurate rate for discrimination was up to 95.00%. The results indicate that dynamic cluster and discriminant analysis can be used to evaluate the pathogenicity differentiation of R. solani AG1 ⅠA.

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Occurrence and Population Dynamics of Chironomid Midges in Early Rice Field LI Zhi-yu,YANG Hong,LAI Feng-xiang,FU Qiang,HU Yang 2010, 24(6): 630-634 .  DOI: 10.3969/j.issn.1001-7216.2010.06.011 Abstract ( )   PDF (3140KB) ( )   The chironomid midge is one of world wide distribution insects and normally occurs in a great abundance among many kinds of water bodies. In rice field, chironomid midges are very abundant. Currently, our knowledge about chironomid midges in rice field is limited to the species survey as a neutral group. By using specially designed emergence traps, the occurrence of chironomid midges in rice field were studied throughout the early rice season in Fuyang, Zhejiang Province, China. Four species of chironomid, namely, Tanytarsus formosanus, Chironomus sinicus, Polypedilum nubifer and Tanypus punctipennis, were collected from rice fields. All of them are widely distributed species. T. formosanus and C. sinicus were the most dominant species in rice field. The dynamic of chironomid midges in early rice field could be divided into two phases, namely early and late phases. The abundance of chironomid midges was higher in the early phase than that in the late phase. It was estimated that there were 45 million chironomid midges emerged from the 1 hectare rice field throughout the early rice season. It was also found that the sex ratio of T. formosanus was female biased during the early phase, whereas it was male biased during the late phase. In contrast, the sex ratio of C. sinicus was male biased throughout the field season.

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Effects of Pymetrozine on the Feeding Behaviour of Rice Brown Planthopper, Nilaparvata lugens (Stal) HE Yue-ping,CHEN Li,CHEN Jian-ming,CHEN Lie-zhong,ZHANG Jue-feng 2010, 24(6): 635-640 .  DOI: 10.3969/j.issn.1001-7216.2010.06.012 Abstract ( )   PDF (2379KB) ( )   Pymetrozine is one of the recommended alternatives for controlling rice brown planthopper, Nilaparvata lugens (Stl). Initial feeding choice experiments with winged female adults showed that after 24 h, there were no significant differences between the numbers of planthoppers alighting on untreated rice plants and those treated with 01 g/L pymetrozine (15±09 adults and 16±12 adults, respectively). Electrical penetration graph (EPG) data indicated that after the treatment of 01 g/L pymetrozine, no significant difference occurred on the frequency of stylet probing, the frequency of penetration into phloem, the duration in the extracellular region of phloem, and the duration of xylem ingestion, while the duration of nonprobing period increased, and the phloem ingestion behaviour was significantly interrupted compared with the treatment of water. The total duration of phloem ingestion of N. lugens adults in 4 h was only 12±05 min on the plants treated with pymetrozine, whereas 651±113 min on the untreated plants. Feeding recovery data suggested that the inhibition of pymetrozine on the phloem ingestion of N. lugens could be recovered, but took long time. The total duration of phloem ingestionof N. lugens was only about 10 min when the insects were first released to the rice seedlings treated by 01 g/L pymetrozine for 24 h, and then survived insects were transferred to the untreated rice seedlings for 120 h. It is revealed that pymetrozine had no deterrent or antifeeding effects on N. lugens, and did not block the stylet probing and the xylem ingestion of N. lugens. However, it significantly inhibited the phloem ingestion of N. lugens, and the recovery of N. lugens from the inhibitive effects was slow. Thus, pymetrozine has a high and persistent efficacy against N. lugens.

综述与专论

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Thoughts and Practice of Some Problems about Research and Application of TwoLine Hybrid Rice CHEN Li-yun,LEI Dong-yang,TANG Wen-bang,XIAO Ying-hui 2010, 24(6): 641-646 .  DOI: 10.3969/j.issn.1001-7216.2010.06.013 Abstract ( )   PDF (32KB) ( )   The main problems about research and application of twoline hybrid rice were reviewed, including the confusional name and classification of male sterile lines, the unclear characterization of responses to photoperiod and temperature and the unsuitable site selection for seed production of male sterile line and hybrid rice. In order to efficiently and accurately use photo(thermo)sensitive genic male sterile (PTGMS), four types, including PTGMS, TGMS, negative PTGMS and negative TGMS were proposed. A new idea for explaining the mechanism of sterility in PTGMS rice was proposed. The transition from sterile to fertile was involved in cooperative regulation of majoreffect sterile genes and the photoperiod and/or temperature sensitive ones for development. The minoreffect genes, with accumulative effect on sterility, were the important factors that affect the critical temperature of sterility transfer. In order to make better use of PTGMS, the characterization of responses to photoperiod and temperature of PTGMS should be made. The identification method for the characterization responding to photoperiod and temperature of PTGMS was also put forward. The optimal ecological site for seed production could be determined according to the historical climate data and the requirements for the meteorological conditions during different periods of seed production.

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Recent Advances in Proteomic Studies on Rice Pathogen Interactions YU Chu-lang,YANG Yong,WANG Xu-ming,YAN Cheng-qi,CHEN Jian-ping* 2010, 24(6): 647-651 .  DOI: 10.3969/j.issn.1001-7216.2010.06.014 Abstract ( )   PDF (38KB) ( )   The molecular mechanisms of plantpathogen interactions have been a longtime hotspot of plant pathology. Recently, functional genomic strategies, including proteomics and transcriptomics, have been widely used in defining gene and protein function and expression profiles. Proteomics, a complementary research content of transcriptomics, has been a powerful tool to systematically analyze the cellular protein expression profiles and posttranslational modifications under pathogen invasion. Rice, a monocotyledonous plant for which the entire genome was completely sequenced, has been adopted as an ideal model for studying the molecular mechanisms of plantpathogen interactions. In this review, the main research techniques of proteomics, recent advances in proteomic analyses of rice under bacteria, fungi and viruses invasion were highlighted, and an overview on phosphoproteomics analyses of ricepathogen interactions was presented.

研究简报

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Gnetic Analysis and Gene Mapping of a Short Root Mutant ksr1 in Rice NING Yong-qiang,DING Wo-na,ZHU Shi-hua*,YU Hong-wei,YU Hong,LU Kai-xing 2010, 24(6): 652-654 .  DOI: 10.3969/j.issn.1001-7216.2010.06.015 Abstract ( )   PDF (1658KB) ( )   A short root mutant ksr1 with the Kasalath background was isolated from an EMSmutagenized population in rice. The root length of 6dayold ksr1 plants was only about 20% of the wild type. Genetic analysis indicated that the short root phenotype of ksr1 was controlled by a recessive mutation in a single nuclearencoded gene. To map the ksr1 mutation, an F2 population was generated by crossing the ksr1 mutant with Nipponbare. The KSR1 locus was linked to the SSR marker RM1223 on chromosome 4. Another eight SSR markers and two InDel markers were developed around this marker. The KSR1 gene was further mapped to a 155 kb region, flanked by the InDel marker 424725K and the SSR marker RM17182.

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Testing Early Generation Stability of a Japonica Rice Variety 84-15 by Using Two Qualitative Traits LU Zuo-mei ,XU Bao-qin,SUN Da-yun 2010, 24(6): 655-658 .  DOI: 10.3969/j.issn.1001-7216.2010.06.016 Abstract ( )   PDF (31KB) ( )   Early generation stability of 8415, a japonica rice variety, was retested by using allelic differences of two qualitative traits. Results showed that both glume tip color and waxy endosperm segregated in progenies of single F1 plants derived from two crosses of 8415/Zi 18 (purple color) and 8415/Dahuaxiangnuo(waxy). The results denied the conclusion obtained in previous studies that 8415 had a characteristic of early generation stability. Meanwhile, it was found that using coefficient of variation as an indicator of early generation stability was unreasonable by analyzing plant height as an example. Importance of avoiding systematic error in research works was also discussed.

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QTL Analysis on Stomatal Trait of Flag Leaf in Rice LI Rui,ZHAO Shu-li,MAO Ting,XU Zheng-jin ,CHEN Wen-fu 2010, 24(6): 659-62 .  DOI: 10.3969/j.issn.1001-7216.2010.06.017 Abstract ( )   PDF (851KB) ( )   To understand the genetic mechanism of stoma related traits, an F10 population derived from a cross between a tropical japonica rice (Akihikari) and an indica rice(Qishanzhan) was used to analyze QTLs controlling the stoma related traits. Nine QTLs for stoma related traits were detected. Four putative QTLs(qSD1, qSD4, qSD7, qSD8) controlling stomatal density were located on chromosomes 1, 4, 7 and 8, with the explained variance of 7.2%, 8.1%, 9.1% and 5.9%, respectively. Two QTLs(qSL4, qSL6) for stomatal length were detected on chromosomes 4 and 6, with the explained variance of 6.6% and 10.5%, respectively. Three QTLs(qSW5, qSW7, qSW10) for stomatal width were detected on chromosomes 5, 7 and 10, with the explained variance of 12.4%, 9.7% and 5.6%, respectively. All the eight QTLs except qSD1 were not previously reported. qSD4 on chromosome 4 controlling stomatal density was overlapped with the QTL of photosynthetic rate which was detected before.

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Frequency of Gene Flow from HerbicideResistant Transgenic Rice to Conventional Rice Varieties LU Yong-liang,PENG Yu-fa,WANG Wei-xia,SUN Xing-qiang,CHEN Li-juan,YU Liu-qing 2010, 24(6): 663-666 .  DOI: 10.3969/j.issn.1001-7216.2010.06.018 Abstract ( )   PDF (461KB) ( )   To observe the frequency of gene flow, a japonica rice 991 that contains the bar gene conferring Basta resistance was used as the pollen donor, and four conventional rice varieties as recipients were planted around it. The results showed that the frequency of transgene flow was 0.0193%-0.3960% at 0-2 m zone, but the frequencies were significantly different in different rice varieties.

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Subspecific Differentiation of Male Sterile Lines of Rice Derived from Indica/Japonica Cross and Heterosis Performance of the F1 Hybrids CHEN Shen-guang,CAO Li-yong,SI Hua-min,CHENG Shi-hua* 2010, 24(6): 667-671 .  DOI: 10.3969/j.issn.1001-7216.2010.06.019 Abstract ( )   PDF (36KB) ( )   Three cytoplasmic male sterile (CMS) lines were developed through the hybridization of indica and japonica rice. 064A and Xieyan A are wild abortivetype CMS lines, and 300S is a photo(thermo)sensitive male sterile line. By using morphological index and RFLP markers, the subspecific differentiation of the male sterile lines were detected. The morphological indexes for 064A, Xieyan A and 300S were 11, 9 and 12, respectively, attributing to the indicalinous type. The analysis of RFLP markers showed that these three lines were clustered into one class, similar to that by morphological index. The results indicate that these male sterile lines have proportional japonica pedigree on their indica genetic background. Some F1 between these three male sterile lines and restorer lines with wide genetic bases performed very strong heterosis in yield and applicable potential in super hybrid rice breeding program.

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Effects of Long-Term Rice-Rice-Green Manure Rotation on Rice Yield GAO Ju-sheng,CAO Wei-dong,DONG Chun-hua,XU Ming-gang,ZENG Xi-bai,WEN Shi-lin, 2010, 24(6): 672-676 .  DOI: 10.3969/j.issn.1001-7216.2010.06.020 Abstract ( )   PDF (4021KB) ( )   In order to study the potential yield of multiple cropping systems in red paddy soil in Southern Hunan Province, continuously improve the economic benefit and soil fertility, promote agricultural sustainable development, a longterm ricericegreen manure rotation experiment has been arranged since 1982. In the experiment, milk vetch(Astragalus sinicus), oilseed rape （Brassica napus) and rye grass (Lolium) were planted as green manures, and represented by T1, T2 and T3, respectively, and winter fallow as CK. Compared with CK, planting green manures could promote rice yield with the order of T1>T2>T3. The difference of annual rice yield between planting green manures and CK increased with the passing years, so did the differences between T1 and T2, T1 and T3, T2 and T3.

实验技术

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A Simple Method for Rapid Preparation of Rice Genomic DNA SUN Chuan,CHEN Gang,RAO Yu-chun,ZHANG Guang-heng,GAO Zhen-yu,LIU Jian,JU Pei-na,HU Jiang,GUO Long-biao,QIAN Qian,ZENG Da-li 2010, 24(6): 677-680 .  DOI: 10.3969/j.issn.1001-7216.2010.06.021 Abstract ( )   PDF (706KB) ( )   A simple method for preparation of rice genomic DNA was developed. A small amount (1- 50 mg) of leaf tissue of rice seedling, 500 μL of extraction buffer, and one steel bead were put into a 2mL microcentrifuge tube. After vigorously mashing for 2 min, 5 μL of the supernatant was directly applied to PCR amplification. Otherwise, the supernatant was precipitated with 2 volumes of ethanol to obtain high quality genomic DNA. This method is simple, rapid, low cost, and reliable for PCR analysis. One person can manipulate as many as 96 samples for PCR in 10 minutes. It is especially suitable for genotyping of large number of samples.