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10 November 2005, Volume 19 Issue 6 Previous Issue    Next Issue
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研究报告
综述与专论
实验技术
研究简报


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研究报告

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Development of the Codominant Marker of Rice Blast Resistance Gene Pi-ta WANG Zhong-hua ,Redus MARC ,JIA Yu-lin 2005, 19(6): 483-488 .  Abstract ( )   PDF (535KB) ( )   A single nucleotide length polymorphism (SNLP) was identified at the intron region of the Pi-ta gene to develop a codominant Pi-ta gene marker suitable for genotyping with an ABI automated machine. The DNA primer specific to the resistance Pi-ta allele was labeled with blue dye as a forward primer, the DNA primer specific to the susceptibility pi-ta allele was labeled with green dye as another forward primer and the DNA primer identical to both Pi-ta/pi-ta alleles was unlabeled as the reverse primer for polymerase chain reaction (PCR). Using these three primers, a 181 bp blue peak in homozygous resistant and a green peak of 183 bp in homozygous susceptible, and both peaks in heterozygous plants were produced by PCR. The utility of marker was verified using 12 plants coming from segregating F 2 population and 15 inbred varieties via dominant markers and pathogenicity test. A codominant Pi-ta marker is thus developed for effective Pi-ta assisted selection for rice disease resistance breeding program.

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Study on Pathogen Resistance of OsWRKY Transcription Factor by dsRNA Interference Method WU Ya-wen ,XIE Ke ,LI Jing ,YU Yong-hong ,HU Guo-cheng ,FU Ya-ping ,SI Hua-min ,DOU Cai-hong ,GUO Ze-jian ,SUN Zong-xiu 2005, 19(6): 489-494 .  Abstract ( )   PDF (581KB) ( )   To study the functions of the WRKY superfamily of transcription factors in rice, a hairpin dsRNA interference vector involving in a sequence of conserved WRKY domain, constituted by about 60 amino acid residues,were constructed. The vector was transformed into the embryos of wild parent Zhonghua 11 by Agrobacterium-mediated co-cultivation method. A total of nine transgenic lines with interference phenotypes were identified from 65 independent transgenic rice plants. PCR and Southern blotting analysis indicated that the foreign DNA had been integrated into rice genome of Zhonghua 11, and most of them were single copy. The three selected interference lines showed more resistant to rice blast (Magnaporthe grisea) and bacterial blight (Xanthomonas oryzae pv. oryzae) than WRKY T-DNA insertion mutant T456 and Zhonghua 11, and T456 was slightly more resistant to these two diseases than Zhanghua 11, suggesting that dsRNA interference was successful and it might have suppressed some OsWRKY transcription factors which were highly associated with regulating pathogen genes.

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Identification of Multiple Alleles at the Wx Locus and Development of Single Segment Substitution Lines in Rice ZENG Rui-zhen ,ZHANG Ze-min ,HE Feng-hua ,XI Zhang-ying ,Akshay TALUKDAR ,SHI Jun-qiong ,QIN Li-jun ,HUANG Chao-feng ,ZHANG Gui-quan 2005, 19(6): 495-500 .  Abstract ( )   PDF (438KB) ( )   The microsatellite markers 484/485 and 484/W2R were used to identify the multiple alleles at the Wx locus in rice germplasm. Fifteen alleles were identified in 278 accessions by the use of microsatellite class and G-T polymorphism. Among them, CT 12-G, CT 15-G, CT 16-G, CT 17-G, CT 18-G and CT 21-G are new ones. Seventy-two single segment substitution lines (SSSLs) carrying different alleles at the Wx locus were developed by the use of Huajingxian 74 with the CT 11-G allele as a recipient and 20 varieties containing 12 alleles at the Wx locus as donors. The estimated length of the substituted segments ranged from 2.2 cM to 77.3 cM with an average of 17.4 cM.

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Molecular Cloning and Characterization of Citrate Synthase Gene in Rice (Oryza sativa) ZHANG Shan-shan ,MING Feng ,LU Qun ,GUO Bin ,SHEN Da-leng 2005, 19(6): 501-505 .  Abstract ( )   PDF (445KB) ( )   The full-length OsCS gene encoding citrate synthase was isolated from rice (Oryza sativa L. subsp.japonica). OsCS is 1470 bp long and encodes a polypeptide with 475 amino acids. Its putative protein sequence is high identical to Daucus carota, Nicotiana tabacum, Beta vulgaris subsp., Arabidopsis thaliana, and Citrus junos (>70%). The deduced amino- terminal sequence of OsCS showes characteristics of mitochondrial targeting signal. Southern blot analysis using ORF of the OsCS as probe indicated that this gene exists in multiple copies in rice genome. The band with predicated size of 59 kD was detected by Western blot after induced by 0.4 mmol/L IPTG.

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SSR Marker-Based Mapping of Fertility-Restoring Genes in Rice Restorer Line Milyang 46 LI Guang-xian ,TU Guo-qing ,ZHANG Ke-qin ,YAO Fang-yin ,ZHUANG Jie-yun 2005, 19(6): 506-510 .  Abstract ( )   PDF (487KB) ( )   A SSR marker based linkage map covering chromosome 10 and the short arms of chromosomes 1 and 11 of rice was constructed and applied for mapping fertility-restoring genes in Zhenshan 97A/(Zhenshan 97B/Milyang 46)F6 population consisting of 704 lines. Four QTLs were detected, among which Rf4 on the long arm of chromosome 10 displayed a major effect, Rf3 on the short arm of chromosome 1 had a considerable effect, and qRf10 and qRf11 in the pericentromeric regions of chromosomes 10 and 11, respectively, had minor effects. It was also found that Rf3, qRf10 and qRf11 contributed to higher spikelet fertility in the presence of the major gene Rf4, whereas qRf10 and qRf11 showed little effect in the presence of both Rf3 and Rf4.

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Genetic Mapping of the Dominant Minute Grain Gene Mi3(t) in Rice (Oryza sativa) LIU Ming-wei ,LIU Yong ,WANG Shi-quan ,DENG Qi-ming ,LI Ping 2005, 19(6): 511-515 .  Abstract ( )   PDF (426KB) ( )   Genetic analysis and gene location were conducted on Y34, a minute grain rice. All grains of F 1 populations derived from Y34 and two long grain rice (Shuhui 881 and Shuhui 527) were all minute grains, and the segregation ratios of the F 2 populations both were fitted to 3∶1, showing that the minute grain trait was controlled by a completely dominant gene. Using the F 2 population from the cross between Shuhui 881 and Y34 and simple sequence repeats markers, a completely dominant gene temporarily named Mi3(t), was located between RM282(the genetic distance was 5.1 cM) and RM6283(the genetic distance was 0.9 cM) on the short arm of chromosome 3.

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Starch Properties of Rice Mutant Enriched with Resistant Starch YANG Chao-zhu ,LI Chun-shou ,SHU Xiao-li ,ZHANG Zhi-zhuan ,ZHANG Lei ,ZHAO Hai-jun ,MA Chuan-xi ,WU Dian-xing 2005, 19(6): 516-520 .  Abstract ( )   PDF (450KB) ( )   The rice mutant enriched with resistant starch (RS), described as RS111, was induced from the elite hybrid rice restorer line R7954. The RS content in the hot cooked rice of the mutant was as high as 7.0%, 2.4 times higher than that in the wild type. Obvious differences in starch granule morphology, curve and parameters of DSC and X-ray diffraction were observed between the mutant and the wild type. The mutant was characterized by relatively identical starch granule size, higher rate of round- and oval-shaped starch granules, lower onset temperature (T O), peak temperature (TP), final temperature (T C) and enthalpy of gelatinization (ΔH GEL). The crude lipid, crude fiber and apparent amylose contents in the mutant were enhanced significantly compared to those in the wild type.

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Comparison on the Characteristics of the Isonuclear Alloplasmic CMS Lines in japonica Rice TANG Shu-zhu ,SUN Ye ,ZHANG Hong-gen ,GU Yan-juan ,LU Ju-fei ,TIAN Shun ,YU Bo ,GU Ming-hong 2005, 19(6): 521-526 .  Abstract ( )   PDF (517KB) ( )   The characteristics of two types of isonuclear alloplasmic CMS lines derived from Suqiu, Liuqianxin were studied in male sterile stability, restorable ability, flowering habit and outcrossing rate. Most pollen granules of BT-type and HL-type CMS lines were stained-abortion, and those of WA-type CMS lines were mainly typical-abortion. Selfed seed-setting rate of BT Suqiu A and BT Liuqianxin A were 0.06% and 3.72%, respectively, the sterility of those CMS lines were the most unstable. The pollen abortive degree of CMS lines affected the length of panicle-neck above the sheath of flag leaf, the more serious abortive degree, the shorter the length of panicle-neck above the sheath of flag leaf. Flowering time and flowering duration of BT-type CMS lines were close to its maintainers. Flowering time of HL-type CMS line was later than that of its maintainers, but flowering duration was the same. Flowering time of WA was distinctly laggar and flowering duration was longer, and flowering was not concentrated. The outcrossing rates of BT- and HL-type CMS lines were higher than that of WA-type CMS line. As to the restorable characteristic, BT-type was the best, HL-type the second, and WA-type the worst.

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Analysis on Combining Ability of Dry Weight and Nitrogen Use Efficiency in Rice PIAO Zhong-ze ,HAN Long-zhi ,KOH Hee-jeong ,ZHANG Jian-ming ,LU Jia-an ,LI Pei-de 2005, 19(6): 527-532 .  Abstract ( )   PDF (550KB) ( )   Combining ability of dry weight and physiological N-use efficiency at different stages was analyzed by using 9×9 diallel crosses with nine different type rice varieties under N-fertilized and N-unfertilized conditions. The additive effects was principal in dry weight and physiological N-use efficiency at maximum tillering stage, dry weight and physiological N-use efficiency at booting stage under N-fertilized and N-unfertilized conditions, physiological N-use efficiency at harvesting stage under N-fertilized conditions, grain yield under N-unfertilized conditions while the non-additive effect was dominant in physiological N-use efficiency at harvesting stage under N-unfertilized conditions, and grain yield under N-fertilized conditions was controlled by additive and non-additive effects. The crosses with larger specific combining ability(SCA) effects did not exhibit high physiological N-use efficiency compared to their parents and were not correlated with crosses with good general combining ability(GCA) certainly. The performance and GCA effect for its parents could not be used to forecast the heterosis in hybrid rice.

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Genotypic Differences on Nitrogen Nutrition Characteristics of Rice Germplasms at Seedling Stage CHENG Jian-feng ,DAI Ting-bo ,CAO Wei-xing ,JIANG Dong 2005, 19(6): 533-538 .  Abstract ( )   PDF (634KB) ( )   Through variation and frequency analysis, the genotypic differences in nitrogen utilization efficiency (NUE), nitrogen absorption efficiency (NAE) and nitrogen utilization efficiency response (NUER) of 88 rice germplasms were studied. NUE, NAE and NUER of the tested rice at seedling stage had notable genotypic discrepancy, and were influenced by nitrogen application. NUE differed significantly under low, medium and high nitrogen, and dropped with the increase of nitrogen application. NAE wasn′t significantly different under low and medium nitrogen, but was notably smaller than that under high nitrogen. NUER between low and high nitrogen was more notably higher than that between low and medium nitrogen or between medium and high nitrogen, and the latter two hadn′t significant discrepancy. The coefficients of variation among NUE, NAE and NUER of rice germplasms at seedling stage were greatly different (above 20%) and the order of them was NUER>NAE>NUE. The genotypic distributions of NUE, NAE and NUER of rice germplasms at seedling stage basically resembled a normal curve, and the order of good-of-fitness was NUE>NAE>NUER.

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Effects of Cultivation Methods on Nitrogen Absorption and Use Efficiency of Upland and Paddy Rice ZHANG Ya-jie ,LIN Qiang-sen ,SUN Bin ,DIAO Guang-hua ,YANG Jian-chang 2005, 19(6): 539-544 .  Abstract ( )   PDF (593KB) ( )   Upland rice cultivar Zhonghan 3(japonica)and paddy rice cultivar Wuxiangjing 99-8 (japonica) were used to study the effects of cultivation methods on nitrogen(N) absorption and use efficiency at later growth stage. The rice grain yield was significantly lower under dry cultivation than under submerged cultivation(SC,control) for Zhonghan 3 and there was no significant difference between dry cultivation with plastic film mulching (PFMC) and MC for Wuxiangjing 99-8,and the yield was the lowest under bare cultivation(BC)among the three cultivation methods for two cultivars. N contents in the plants under PFMC and BC were lower at heading,and decreased more slowly from heading to maturity with the order of SC > BC >PFMC. The amount of N absorption by plants was in the order of SC > PFMC > BC at heading and maturity. The proportion of N was significantly higher in the culms and sheaths,whereas significantly lower in the grains,under dry cultivation than under SC. The proportion of N in leaves was different between the two cultivars. N efficiency of matter production was significantly greater under dry cultivation than under SC at heading and was the least for PFMC among the three cultivation methods at maturity. Dry cultivation had higher N efficiency of grain yield production(except PFMC for Zhonghan 3)and greater N harvest index than SC,with the order of BC > PFMC > SC. Compared with Wuxiangjing 99-8,Zhonghan 3 exhibited a faster decrease in N content after heading,smaller N accumulation in plants,greater proportion of N in leaves and grains and smaller in the culms and sheaths,higher N efficiency of matter production(at maturity) and grain yield production,greater N harvest index,and lower grain yield.

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Effects of Weak Light on Rice Starch Accumulation and Starch Synthesis Enzyme Activities at Grain Filling Stage LI Tian ,Ryu OHSUGI ,Tohru YAMAGISHI ,Haruto SASAKI 2005, 19(6): 545-550 .  Abstract ( )   PDF (575KB) ( )   Dynamic changes of starch content, amylose content, sucrose content and the activities of starch synthesis enzymes under shading treatment after flowering were studied using IR72 (indica) and Nipponbare (japonica) as material. Under shading treatments, starch, amylose and sucrose contents in the rice grain decreased, while ADP-glucose pyrophosphorylase (ADPGPPase) activity only changed a little, soluble starch synthase activity and granule bound starch synthase activity were decreased, soluble starch branching enzyme activity and granule bound starch branching enzyme activity were increased. Starch debranching enzyme activity was decreased in IR72, while increased in Nipponbare compared with the normal treatment. Correlation analysis showed that changes of starch content were positively and significantly correlated with changes of sucrose content under weak light. Both ADPGPPase activity and soluble starch branching enzyme activity were positively and significantly correlated with starch accumulation rate. It was implied that the decline of starch synthesis enzyme activities was relative to the decrease of starch component, and the increase of the activity of starch branching enzyme played an important role in the decrease of ratio of amylose to the total starch under weak light.

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Isolation and Identification of Allelochemicals from Wheat and Allelopathy on Leptochloa chinensis in Direct-Seeding Rice Field DONG Li-yao ,WANG Ming-hua ,WU Shu-wen ,SHEN Jin-liang 2005, 19(6): 551-555 .  Abstract ( )   PDF (490KB) ( )   Allelopathy effects of aqueous extracts from culms and roots of wheat on Leptochloa chinensis (L.) Nees in direct-seeding rice field was investigated and allelochemicals were isolated and identified. The bioassay results showed that aqueous extracts from different varieties of wheat had no allelopathy effects on rice seedling growth, but on seed germination of L. chinensis, which were safe to the seed germination and seedling growth of rice. Allelochemicals from wheat, which could inhibit the seed germination of L. chinensis, were isolated and identified by use of gas chromatography mass spectrometry (GC-MS) method. It was suggested that organic acid, which were composed of oleic acid, palmific acid, myristic acid, linoleic acid, stearic acid, exhibited great allelopthic potential.

综述与专论

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Achievement and Prospects of Research on Evaluation of the Relationship Between Source and Sink in Rice WANG Feng ,ZHANG Guo-ping ,BAI Pu 2005, 19(6): 556-560 .  Abstract ( )   PDF (602KB) ( )   Rice yield is mainly dependent on the strength of source, sink, stream and their harmonization. There have been many reports on evaluation of relationship between source and sink, but no consistent criteria are commonly recognized. Advances of research on evaluation of source-sink relationship in rice was reviewed in terms of the content and criteria of source, stream, and sink, and interactions among these three factors, and their effects on grain development. Meanwhile, the various methodologies for evaluating the source-sink relationship and classifying rice varieties being done in China were compared. Finally, the main issues to be done in developing an ideal evaluation system of source-sink relationship were also proposed.

实验技术

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A Simplified Rice DNA Extraction Protocol for PCR Analysis CHEN Wen-yue ,BAO Jin-song ,ZHOU Xiang-sheng ,SHU Qing-yao 2005, 19(6): 561-563 .  Abstract ( )   PDF (320KB) ( )   A simple protocol was established for DNA extraction by using rice etiolated seedlings. By using this method, rice DNA was directly extracted in 0.5 mol/L NaOH solution in an eppendorf tube. Results of comparative PCR analyses and electrophoresis showed that DNA extracted by using this method was as good and useful as that isolated by using standard CTAB method.

研究简报

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Development of Amylose Determination System Based on Computerized Colorimetric Measurement(CCMA) CHANG Guo-hua ,HOU Cai-yun ,SHANG Yan-fen ,CUI Gai-beng ,ZHOU Xiao-feng 2005, 19(6): 564-566 .  Abstract ( )   PDF (294KB) ( )   A system based on the computerized colorimetric measurement(CCMA) method was developed by using the Microsoft C ++. It could determine the content of amylose in rice, and give out the quality grade automatically according to the standard performed. The system would be available to determine the chemical quality quickly and efficiently by using image processing technology.

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Effects of Water-Saving and Protective Tillage Technique in Rice LI Jin-feng ,XU Chun-lin ,CHU Jiang ,LU Yan-dong ,LI Hong-yu 2005, 19(6): 567-569 .  Abstract ( )   PDF (296KB) ( )   The soil and rice plant characteristics, and grain yield and its components of rice under water-saving and protective tillage (WSPT) conditions were analyzed. Compared with conventional tillage method, the soil bulk density were decreased, and soil porosity, redox potential, available N, P, K contents in the soil layers of seedling were increased, the root dry weight and leaf N content of rice were enhanced under WSPT. The yield under WSPT was attributed to the increase in number of panicles per unit area and 1000-grain weight.

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Effect of Saline-Alkali Soil from Songnen Plain on Chlorophyll Content in Rice SHAO Xi-wen ,ZHANG Rui-zhen ,TONG Shu-yuan ,DU Zhen-yu ,YANG Mo ,SUN Chang-zhan 2005, 19(6): 570-572 .  Abstract ( )   PDF (273KB) ( )   Effect of different saline-alkali stress on chlorophyll, chlorophyll a and chlorophyll b contents of two rice cultivars (Ji 89-45 and Jinongda 10) were studied with pot experiments. The chlorphyll contents were low at the early growth stage,then increased gradually and reached the highest value at the flowering and milk stage, and no difference in change tendency of chlorphyll content between the two cultivars was found. Chlorphyll contents under the stress treatment were lower than those under control treatment. In addition, the variable range of chlorphyll a content was always wider than that of chlorophyll b content.

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Resistance Screening and Evaluation of Newly-Bred Rice Varieties (Lines) to the Rice Brown Planthopper, Nilaparvata lugens CHEN Jian-ming ,YU Xiao-ping ,CHENG Jia-an ,LU Zhong-xian ,ZHENG Xu-song ,XU Hong-xing 2005, 19(6): 573-576 .  Abstract ( )   PDF (428KB) ( )   Resistance of 769 newly-bred rice varieties or lines(1999-2002) to the brown planthopper (BPH) (Nilaparvata lugens) was evaluated using the Modified Seedbox Screening Technique (MSST). Among the tested rice varieties, 93 varieties had resistance to BPH, of which 3, 13, 77 varieties had long-, middle- and short durable resistance respectively at seedling stage as TN1(check) was graded 9. Meanwhile, the resistance levels of the 47 resistant varieties tested were further evaluated at adult stage. There were 3, 3, 15 varieties expressed long-, middle- and short-durable resistance at adult stage respectively with the same tendency as those at seedling stage. Twenty-four short-durable (SD) resistant varieties at seedling stage had three middle-durable (MD) and 21 no durable resistance at adult stage respectively, and three MD resistance varieties at seedling stage expressed SD resistance at adult stage. It is an effective method for evaluating tolerance levels of rice varieties to BPH usingonclick="SelectDisplayDiv('EnDivSummaryMoreSummary','EnDivSummaryMore');DisplaySpanDiv('EnDivSummaryHuanYuan')"> damage (grade 5) and durable resistance level (MD to LD) at seedling and adult stages.

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Primary Study on Resistance of Rice Stem Borer (Chilo suppressalis) to Fipronil JIANG Wei-hua ,HAN Zhao-jun ,HAO Ming-li 2005, 19(6): 577-579 .  Abstract ( )   PDF (337KB) ( )   By means of topical application, fipronil resistance was surveyed in the rice stem borer, Chilo suppressalis,from 12 diferent areas in East China from 2001 to 2004. The rice stem borers in most regions of Jiangsu and Anhui were still susceptible to fipronil. But in Wuxi of Jiangsu and Cixi of Zhejiang, their sensitivity decreased (resistance ratios were 3.1 and 3.6), and the moderate level of resistance (resistance ratio was 21.2) was found in the population of Cangnan, Zhejiang. Therefore it was concluded that the pest resistant to fipronil was still at the early developmental stage. Synergism experiments showed that PBO had little effect on both susceptible and resistant borers (synergism ratios were 1.1 to 1.2). Though TPP and DEM had no effect on susceptible ones, they had significant synergism to fipronil in resistant borers (synergism ratios were 1.8 and 16, respectively), indicating esterase and GSTs might be involved in the resistance mechanism. Bioassay with currently used insecticides revealed that triazophos (because of high resistance), trichlorphon and acephate had very low toxicity to resistant borers, while diazinon, pyridaphenthion, decamethrin and avermectin showed high toxicity and had no cross resistance to fipronil, which could be considered as substitute insecticides in the resistance managment.