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10 May 2004, Volume 18 Issue 3 Previous Issue    Next Issue
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研究报告
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Developing Putative Rice SNPs Through Bioinformatics Approaches YANG Lun#,SHEN Wen-biao,CHEN Hong,JIANG Yang,ZHANG Lin,XU Lang-lai * 2004, 18(3): 185-191 .  Abstract ( )   PDF (296KB) ( )   The method and the corresponding progress in developing single nucleotide polymorphism (SNP) of human genome were summarized firstly, and then the latest research progress in rice SNPs related to grain quality, resistance or tolerance to different biotic or abiotic stress was presented. Combined with authors′ latest research progress, including homeodomain leucine-zipper transcriptional factor encoding genes, the availability of developing putative rice SNPs through bioinformatics approaches was surveyed. The importance of developing rice SNPs for the rice breeding and functional genomics research was also elucidated.

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Monitoring and Analyzing the Genes Specific-Expressed at Flowering Stage of Rice (Oryza sativa) WANG Li-hua ,DENG Ye ,FANG Yong-qi ,TAN Xue-lin ,LI De-bao 2004, 18(3): 192-198 .  Abstract ( )   PDF (297KB) ( )   A 2200 uni-gene cDNA array was hybridized with the total mRNA from rice at flowering stage for monitoring the expression profile. Results showed that 356 genes were up-regulated(20.70%), 472 genes were down-regulated(27 44%) and 892 genes were housekeeping(51.86%). Many genes expressed during the flowering process and embryo development such as putative peptide methionine sulfoxide reductase, chalcone synthase, acyl carrier protein Ⅱ gene, gene for allergenic protein, prolamine, 3-ketoacyl carrier protein synthase Ⅲ, and beta-d-glucan exohydrolase-like protein. In order to confer these genes transcription on flowering, mRNA for chalcone synthase, acyl carrier protein Ⅱ gene, prolamine, methionine sulfoxide reductase and gene for allergenic protein were chosen to hybridize with a 755-RNA (bio-phenotypes) array. Results verified that prolamine and allergenic protein were high-expressed in filling grains after being regulated by development, chalcone synthase and methionine sulfoxide reductase were high-expressed in the leaves induced by light, in the root stressed for nitrogen deficiency. Acyl carrier protein Ⅱ was a house-kept gene at all the phases except for at seedling with significant down-regulated characteristics. A co-regulation existed in the expression among chalcone synthase, methionine sulfoxide reductase and acyl carrier protein Ⅱ, and so did allergenic protein and prolamine.

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Analysis of Nucleotide Binding Proteins and Nucleic Acid Binding Proteins of Rice Gall Dwarf Virus and Rice Dwarf Virus and Their Binding Ability LIANG Jian-she,ZHONG Bo-xiong 2004, 18(3): 199-202 .  Abstract ( )   PDF (1457KB) ( )   By incubation of virion with 〔α- 32P〕GTP, 〔α- 32P〕ATP, 〔α- 32P〕CTP and 〔α- 32P〕UTP and Northwestern blotting, the function of structural proteins from rice gall dwarf virus (RGDV) was determined. In the structural proteins, the P1 was the putative RNA-dependent RNA polymerase, which might play an important role in the synthesis of RGDV genomic dsRNAs;The P5 bound with GTP and UTP was a NTP binding protein with RNA guanylyltransferase activity that synthesized a cap structure (mGpppAmpGp) for mRNA of RGDV; The P6 was RNA binding protein and might play an important role in RGDV particle package, as a “key protein” for the assembly of genomic RGDV dsRNAs into the viral particle. The structural protein with the strongest binding ability to nucleic acid was the nucleic acid binding protein of the protein P7 in rice dwarf virus (RDV) which has the similar biological function with the structural protein of the protein P6 in RGDV; the second one was RNA-dependent RNA polymerase; and the lowest one was RNA guanylyltransferase. It indicated that the biological function of the three structural proteins of RDV or RGDV could be judged auxiliary by their binding ability to the nucleic acid.

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Study on the Function of OrLPT1, a Possible Gene Encoding High Affinity Phosphate Transporter in Rice (Oryza sativa) MING Feng ,LU Qun ,DAI Wei ,SHEN Da-leng , 2004, 18(3): 203-207 .  Abstract ( )   PDF (808KB) ( )   Based on the cDNA attained before, the full-length OrLPT 1 was completely sequenced by the RACE. OrLPT 1 is 1745-bp long, encoding a 535 amino acids polypeptide, which has conserved domain of N-linked glycosylation, casein kinaseⅡand protein kinase C. The OrLPT 1 plays an important role in increasing phosphorus uptake ability of rice root under those of low-phosphorus stress. The uptake rate and phosphorus amount of transferred calli are both higher than those of non-transferred calli.

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A Toxicity Test of Transgenic Rice with mtlD/gutD Gene On Rats WANG Hui-zhong ,YING Qi-cai ,ZHONG Guo-qing ,TIAN Ju-xia ,CHEN He,HUANG Da-nian ,QIAN Qian ,ZHANG Hong-sheng 2004, 18(3): 208-212 .  Abstract ( )   PDF (189KB) ( )   Toxicity experiments, mutation experiments and 30-day feeding tests were conducted on rats with transgenic rice bearing mtlD/gutD gene. Results showed that oral LD 50 for rats was larger than 30 g/kg and no mutations were found. The rats groups with 18,36,54 g/kg-dosage had good development in 30-day feeding test. No obvious differences were found in the index of weight, food utilization rate, routine blood test, rate of organ to body (weight to weight), and patho-histological observations between the rats fed with or without the transgenic rice grain. It could be concluded that transgenic rice with mtlD/gutD gene was safe to the rats.

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Analysis on a New Genetic Behavior of Elongated Uppermost Internode in Rice ZHANG Yi,HE Guang-hua *,YANG Guang-wei *,XIANG Juan,SHEN Fu-cheng,YANG Zheng-lin 2004, 18(3): 213-217 .  Abstract ( )   PDF (183KB) ( )   An eui(elongated uppermost internode) mutant was found from wild type of Luoshami, in which the uppermost internodes contribute to the variation of plant height. The eui mutation also resulted in significant increase in flag leaf and grain length. The crosses from mutant type/wild type, wild type/mutant type, mutant type/Jinhui 10 and mutant type/Jin 1B expressed normal uppermost internodes in F1, and segregated differently in F2(ratio of normal to eui was 3∶1 in F2 between wild and mutant type, and 13∶3 in F2 between mutant type and Jinhui 10, Jin 1B). Inheritance analysis indicated that the eui character is controlled by not only a major effective gene but also an inhibited gene. Two genetic systems were proposed to explain the hereditary behaviors. One was that the eui character is controlled by a dominant major gene of Eui and the Eui gene is inhibited by a dominant gene I, and the other was the eui character is controlled by a recessive major gene of eui and the eui gene is inhibited by a recessive gene i.

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Methods of Constructing Core Collections for Rice Germplasm by Using the Genotypic Value LI Chang-tao,SHI Chun-hai*,WU Jian-guo,XU Hai-ming,ZHANG Hai-zhen,REN Yu-ling,FEI Wan-xin 2004, 18(3): 218-222 .  Abstract ( )   PDF (198KB) ( )   Methods of constructing core collections for rice were used to evaluate 992 rice varieties with 13 quantitative traits. The genotypic values of the varieties for the traits studied were predicted by using the adjusted unbiased prediction (AUP) method. Based on the genotypic values, the Mahalanobis distances were calculated and employed to measure the genetic similarities among rice varieties. Three hierarchical clustering methods, the complete linkage method, the Ward’s method and the unweighted pair-group average method, combining with three sampling strategies including the random sampling, the preferred sampling and the deviation sampling, were proposed to develop nine core collections of rice germplasm. Results showed that larger genetic variation of the initial collection could be retained by using the deviation sampling method along with the unweighted pair-group average method of hierarchical clustering. The core collections based on genotypic values had more genetic diversities than those of phenotypic values.

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Dry Matter Production and Translocation and Yield Formation Characters of Different Two-Line Hybrid Rice Combinations with the Same Restorer Line TU Nai-mei,ZHOU Wen-xin,ZHENG Hua,LIU Ai-yu 2004, 18(3): 223-228 .  Abstract ( )   PDF (259KB) ( )   Three two-line hybrid rice combinations and their same restorer line were used to study their dry matter production and translocation characters. The leaf area index and sucrose-phosphate synthase activity of hybrid rice combinations were higher than those of their restorer line at heading stage, but no difference existed after then. The net assimulate rate and crop growth rate of hybrid rice combinations were higher than or similar to those of restorer line from booting to heading. The products of the ratio of saccharide in stem and sheath at heading stage and spikelet weight, and translocation rate of saccharide in stem and sheath were lower in three hybrid rice combinations than those in restorer line. Transport characters of restorer line were supperior to those of hybrid combinations; but the catalase activity in upmost internode of restorer line was higher than that of hybrid combinations. The seed setting rate of hybrid combinations was lower than that of restorer line. The yield of Lu 18S/1125 was significantly higher than that of restorer line, and no significant differences between other two combinations and restorer line were observed.

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Effects of Naphthylacetic Acid and Uniconazole on Root Physiological Characteristics on Super Japonica Hybrid Rice Liaoyou 3225 HUA Ze-tian ,CHEN Wen-fu ,WANG Yan-rong ,DAI Gui-jin ,HAO Xian-bin ,ZHANG Zhong-xu ,WANG Yan ,LI Quan-ying 2004, 18(3): 229-233 .  Abstract ( )   PDF (222KB) ( )   When rice roots were soaked with naphthylacetic acid(NAA) and uniconazole before transplanting or sprayed on plants before heading, rice root growth, root vigor and function were improved at grain-filling and ripening stages, and root senescence was delayed. NAA had more effect on accelerating root growth at early stage, and making root activity decline quickly at late ripening stage; Uniconazole made root grow more evenly in the whole growth period, namely moderately weakened root growth at early stage and strengthened at late stage. NAA and uniconazole sprayed on plant before heading were more effective on improving root growth and activity at late stage than soaking root before transplanting.

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Effect of Exogenous Sorbitol on Photosynthetic Characteristics and Polypeptide Compositions of Thylakoid Membrane of Liangyoupeijiu and Wuyunjing 7 under Salt Stress YANG Yan-hua ,CHEN Guo-xiang ,LIU Shao-hua ,ZHOU Quan-cheng ,CHEN Li ,WANG Gui-min ,LU Chuan-gen 2004, 18(3): 234-238 .  Abstract ( )   PDF (774KB) ( )   Photosynthetic characteristics and polypeptide compositions of thylakoid membrane in four-leaf-stage rice seedlings of Liangyoupeijiu and Wuyunjing 7 under salt stress were studied when rice plants were sprayed with sorbitol. With increase of salt concentration, chlorophyll content, electron transport activity of PSⅡ, the O2 evolution of chloroplast and photosynthetic rate decreased, the ATP content in chloroplast, ATPase activity of chloroplast first boosted up and then dropped, but electron transport activity of PSⅠ, respiration rate enhanced. There were not obvious differences on the polypeptide compositions of thylakoid membrane. The resistance to salt stress in Liangyoupeijiu is obviously stronger than that in Wuyunjing 7. Exogenous sorbitol has some mitigative functions to salt stress.

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Effects of Cadmium Stress on Plant Growth and Antioxidative Enzyme System in Different Rice Genotypes SHAO Guo-sheng ,MUHAMMAD Jaffar Hassan ,ZHANG Xiu-fu ,ZHANG Guo-ping 2004, 18(3): 239-244 .  Abstract ( )   PDF (220KB) ( )   The growth, lipid peroxidation and antioxidant enzyme system of two rice cultivars (Bing 97252, low grain Cd accumulation, and Xiushui 63, high grain Cd accumulation) were studied with a hydroponic experiment under four Cd levels (0.0, 0.1, 1.0 and 5.0 μmol/L). Cd stress reduced plant height and chlorophyll content in rice leaf, altered malondialdehyde (MDA) content and activities of superoxide dismutase (SOD), catalase(CAT) and peroxidase (POD) in rice leaf. Roots and shoots of rice plants responded differently to Cd stress in terms of antioxidant enzyme activity. Generally, the activities of SOD, POD and CAT in shoots and roots dropped with increase of Cd level, while MDA content increased with Cd level, which would be more pronounced in Xiushui 63 than Bing 97252. At booting stage a decrease of 46-52% in SOD activity was noted in the roots of plants exposed to 5.0 μmol/L Cd, while at tillering stage the decrease was 13-19% compared to the control. A significant decrease in chlorophyll content in rice leaf and plant height was noted in higher Cd treatment (1.0 and 5.0 μmol/L). The higher MDA and less chlorophyll content were found in Xiushui 63, indicating that Xiushui 63 was more sensitive to Cd stress than Bing 97252.

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Effects of Indole Acetic Acid(IAA) on the Negative Phototropism of Rice Root MO Yi-wei,WANG Zhong*,QIAN Shan-qin,GU Yun-jie 2004, 18(3): 245-248 .  Abstract ( )   PDF (929KB) ( )   To properly explore the effects of IAA on the negative phototropism of rice(Oryza sativa L.)root,immunoassay of ELISA(enzyme linked immunosorbent assay) was employed to determine IAA concentration in root tip, unilateral application of agar block containing IAA on root tip was carried out to investigate the effect of exogenous IAA on phototropic response. And the bending part of the rice root was sliced into semi-thin sections to study its microstructure. Results showed that the growth of seminal roots could be regulated by exogenous IAA as well as light, the root was induced to bend toward the site of the application, caused asymmetric growth of the root cells at the elongation zone and resulted in the bending growth. IAA concentration on the shaded side of adventitions root increased much greater 1.5 h after the start of irradiation. The unequal lateral IAA distribution can be concluded to be the main cause for the negative phototropism of rice root.

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The Primary Study on Floret and Grain Development Simulation in Rice LIU Xin-wei,MENG Ya-li,ZHOU Zhi-guo,CAO Wei-xing 2004, 18(3): 249-254 .  Abstract ( )   PDF (238KB) ( )   A process-based simulation model for predicting initiated and degenerated floret numbers, grain number and filling dynamics in rice was developed on the basis of published results and field experiments. The model quantified the relationships of panicle organ developmentto physiological development time and environment factors, and non-uniformity of grains on a rice panicle to anthesis timing and assimilate supply, and further simulated the dynamic changes of grain filling at different panicle positions as superior and inferior grains. Testing of the model indicated that the model could effectivelypredict the dynamics of floret development and grain filling in rice.

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Degradation of Cry1Ab Toxin Protein Expressed by Bt Transgenic Rice in Paddy Soils BAI Yao-yu ,JIANG Ming-xing ,CHENG Jia-an ,SHEN Hui-mei ,YANG Pu ,CHEN Zheng-xian ,JIANG Yong-hou ,SHU Qing-yao 2004, 18(3): 255-261 .  Abstract ( )   PDF (278KB) ( )   Two transgenic rice lines, KMD2 and Huachi B 6, both contain a synthetic cry1Ab gene from Bacillus thuringiensis (Bt) under the control of maize ubiquitin promoter, were used in the laboratory to examine the degradation of Cry1Ab toxin proteins from Bt transgenic rice in paddy soils. Ground leaves of KMD2 and whole leaf-blades of Huachi B6 were separately buried in each of three types of soil, i. e. pale paddy soil on quaternary red soil, blue clayey paddy soil and marine-fluvigenic yellow loamy paddy soil, and thereafter residues of Cry1Ab in the KMD2-leaf-mixed soils and Cry1Ab in the buried Huachi B 6 leaves were examined at an interval of 6-10 days. Cry1Ab in ground leaves of KMD2 degraded rapidly in all the soils during 36 days after treatment (DAT), particularly during the first six DAT, with the highest degradation rate in blue clayey paddy soil and the lowest in marine-fluvigenic yellow loamy paddy soil. During 6-36 DAT, the highest Cry1Ab residue was observed in marine-fluvig enic yellow loamy paddy soil, mediate in pale paddy soil on quaternary red soil, and the lowest in blue clayey paddy soil, with significant difference tested among soils at each of observation times; however, such difference decreased gradually as time proceeded thereafter, and became fairly little at 78 DAT. As the soil was flooded, the degradation of Cry1Ab in buried leaves of Huachi B 6 was significantly accelerated, and exhibited a similar pattern among all of the flooded soils. Such impacts of flooding occurred only in the first 12 DAT, and during most of the subsequent periods no significant difference could be observed in Cry1Ab residue between the flooded and unflooded treatments. Degradation kinetics of Cry1Ab toxin proteins were fitted to exponent equation in each treatment, and half life of the toxin protein was determined. Finally, impacts of micro-organism and organic matter content in soils on the degradation of Cry1Ab were discussed.

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The Perfect Stage and SSR Analysis of Magnaporthe grisea in the Yanxi Blast Nursery, Hunan Province SHEN Ying ,Julien FROUIN ,HE Yue-qiu ,Claudia KAYE ,XIAO Fang-hua ,Jean Loup NOTTEGHEM ,LIU Er-ming ,Didier THARREAU 2004, 18(3): 262-268 .  Abstract ( )   PDF (368KB) ( )   One hundred and twenty-four Magnaporthe grisea isolates collected in four years from the Yanxi Blast Nursery, Hunan Province were tested for mating type with four hermaphroditic fertile isolates MAT1.1 (KA3 and TH12) and MAT1 2 (Guy11 and TH16) and the genetic diversity of 105 isolates were analyzed by using microsatellite markers(SSR). Among the 124 isolates, 28 belonged to MAT1.1, 39 to MAT1.2 and the remaining 57 to sterile ones, taking 22.58%, 31.45% and 45 97% for each group, respectively. The 105 isolates could be divided into six lineages at 0.67 similarity level based on the amplicons with seven pairs of SSR primers.

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Frequency and Distribution of Microsatellites in the Whole Genome of Rice Blast Fungus, Magnaporthe grisea LI Cheng-yun,LI Jin-bin,ZHOU Xiao-gang,DONG Ai-rong,XU Ming-hui 2004, 18(3): 269-273 .  Abstract ( )   PDF (194KB) ( )   A total of 37.89 Mb of publicly available DNA sequence in rice blast fungus, Magnaporthe grisea was studied for mono- to hexa-nucleotide simple sequence repeat (SSR or microsatellite) to determine the type, size and frequency. A total of 16 398 SSRs were observed in the whole genomic DNA sequence with criteria of SSR length >15 bp and 80% matches. Considering all six classes of SSR, they occurred on an average about once every 2.31 kb for mono- to hexa-nucleotide in genomic DNA. The most abundant microsatellite was mononucleotide repeat, with the SSR number being 4 392, followed by trinucleotide and pentanucleotide repeats, being 3 586 and 3 442, respectively. These three classes of SSR occupied 69.7% of total microsatellite repeats. The least abundance was dinucleotide repeat, only 680 tracts were found. Repeats of A,AG,AC,ACG,AGC,AAG,GGC,ACCT,ATCC,AAAG,AAAAG,AAAAT,AAAAC,AAAAAG, AAAAAT and AACTAG were predominant, whereas some motifs absent in whole genomic sequence. Comparison of distribution and abundance between different superlinkage groups showed that there was little difference among supperlinkage groups at this level. Availability of these data should facilitate the utilities of SSR on molecular marker developing, population, structural and functional study of non-coding DNA sequence of the phytopathogenic fungus.

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Genetic Expression of Rice Male Sterility in the Allocytoplasmic Background WANG Nai-yuan ,LI Yu ,LIANG Kang-jing ,CAI Zhi-jun ,YANG Ren-cui 2004, 18(3): 274-276 .  Abstract ( )   PDF (143KB) ( )   By using F1 generation of the natural male sterile Jinzao 6/normal fertile Jinzao 6 as male parent, and the same nucleus alien cytoplasmic line developed by hybridization and numerous backcrosses of nine rice germplasms /Zhenshan 97 as female parent, the genetic expression of male sterility was observed. The nine alien cytoplasms included O.barthii,O.glaberrima, Indian T141, African cultivated rice Gambiaka Kokum, Chinese indica rice Zhenshan 97, Indonesian javanica rice Boak and Gannal, Chinese japonica rice Huakenuo and Japanese japonica rice Nanjing 56.Results from F2 generation showed that male sterility of Jinzao 6 segregated and male sterile plants were obtained under all the backgrounds of the nine alien cytoplasms, and no fertile cytoplasmic type caused by interactive inheritance with the male sterile gene of Jinzao 6 was found.

研究简报

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Population Structure of Magnaporthe grisea from Rice in China ZHOU Yi-jun,BAI Juan,CHENG Zhao-bang,FAN Yong-jian,XIONG Ru-yi,WANG Yue 2004, 18(3): 277-280 .  Abstract ( )   PDF (168KB) ( )   Three hundred and twenty four Magnaporthe grisea isolates from rice in China were analyzed by rep-PCR for genetic diversity. DNA fingerprint patterns of the isolates showed that there were many differences among the isolates from different areas. In 20 genetic lineages of M. grisea, CL20 is dominant one in China. All isolates could be classfied into two groups by DNA fingerprint patterns. CN stands for M. grisea from Jiangsu, Zhejiang and North China; CS stands for isolates from Guangdong, Sichuan, etc. Result showed that variation of M. grisea from rice in China is related with variety distribution and climate conditions.