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10 September 2012, Volume 26 Issue 5 Previous Issue    Next Issue
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Reviews and Special Topics

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Research Progress and Breeding Application of Recessive Genic Male Sterility Genes in Rice MA Xiqing1,2, FANG Caichen1,2, DENG Lianwu1,2, WAN Xiangyuan1,2,* 2012, 26(5): 511-520.  DOI: 10.3969/j.issn.10017216.2012.05.001 Abstract ( )   PDF (634KB) ( )   The recessive genic male sterility (RGMS) materials play an important role in hybrid breeding and rice production. However, these materials have not been fully utilized because of unavailable strategies and technology systems to maintain and reproduce these lines. The rapid development of molecular and biological technology provides opportunities to utilize RGMS genes. This review focuses on the research advances of rice RGMS genes and summarizes those RGMS genes in maize, tomato and Arabidopsis. Hopefully, we can obtain more rice RGMS genes by using bioinformatics and new biotechnology methods (e.g., targeted mutation), and can resolve the maintenance and reproduction of RGMS lines effectively and establish a new system for hybrid breeding to enhance utilization of hybridization by combining seed production technology（SPT） with traditional breeding technology in rice.

Research Papers

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Differential Regulatory Mechanism of CBF Regulon Between Nipponbare (japonica)  and 9311(indica) During Cold Acclimation PAN Xiaowu， LI Yongchao， LI Xiaoxiang*， LIU Wenqiang， MIN Jun， LU Tingting， SHENG Xinnian，TAN Jiang 2012, 26(5): 521-528.  DOI: 10.3969/j.issn.10017216.2012.05.002 Abstract ( )   PDF (2394KB) ( )   Nine CBF/DREB1 homology genes in rice were obtained by BLAST search in the NCBI database, which shares conserved amino acid sequences with DREB1 protein in Arabidopsis.  Three CBF  genes organized in tandem, named OsCBF1, OsCBF2, OsCBF3, showed a transient induction in the process of cold acclimation, but much stronger in 9311 compared with that in Nipponbare.  The candidate downstream genes OsLIP5 and OsLIP9  were induced in 9311 but not in Nipponbare.  However, the expression of OsP5CS was induced in both varieties. The differential expression of CBF regulon might be caused by polymorphisms within promoter sequences between these two rice varieties.

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Differential Protein Expression  Analyses of  Pollens of  Honglian CMS Rice at Uninucleate Stage WEN Li1, 3,*, LIU Gai2, WANG Kun2, TAN Bijun1, TAO Jun1 2012, 26(5): 529-536.  DOI: 10.3969/j.issn.10017216.2012.05.003 Abstract ( )   PDF (1749KB) ( )   To better understand the molecular mechanism of cytoplasmic male sterility (CMS) at the protein level during the development of microspores,   the protein expression profile  was analyzed in pollens of the CMS rice line  Yuetai A, its fertile maintainer line Yuetai B and their hybrid F1 (Honglianyou 6). The proteins were separated by two dimensional electrophesis  with immobilized pH (3-10 nonlinear, NL) gradients as the first dimension and SDSPAGE as the second, and 15 differentially expressed proteins were identified by Matrix assisted laserdesorption/ionizaton time of flight mass spectrometry (MALDITOF/MS) or liquid chromatographytandem mass spectrometry (LCMS/MS) analyses. Among these proteins,  some involved in substance and energy metabolism, cell cycle, transcription and cellular transporters etc. were downregulated or absent in the malesterile line, including a putative potassium/proton antiporterlike protein, a zinc knuckle domainlike, a WDrepeat protein etc.  It seemed  to suggest that the downregulation  or absence of these proteins in pollen might relate to the pollen sterility caused by the insufficient mitochondrial energy.

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Screening of Rice Genes Interacted with p5b of Rice blackstreaked dwarf virus LU Ying1,2, YANG Jian2, ZHANG Hengmu2, CHEN Jianping2,* 2012, 26(5): 537-541.  DOI: 10.3969/j.issn.10017216.2012.05.004 Abstract ( )   PDF (741KB) ( )   The second open reading frame on the genome segment 5 (S5) of Rice blackstreaked dwarf virus (RBSDV) encodes a viral nonstructural protein (p5b) with unknown function. To reveal the function of p5b, its gene was ligated into bait plasmid pGBKT7 and an expression library containing rice cDNAs was constructed using plasmid pGADT7 of yeast twohybrid. The bait protein p5b was detected in yeast by western blot and the result of an autoactivation test showed that p5b could not autonomously activate the expression of reporter genes in yeast. Then the bait protein p5b was used for screening the expression cDNA library of rice plant. Gene fragments of some pivotal enzymes, involved in photosynthesis, respiration, and other important metabolism processes, were identified to interact with p5b in yeast, suggesting that these interactions may play roles in symptom development of infected plants.

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Mining Elite Alleles for Growth Duration and Productive Panicle Number per Plant by Association Mapping with Conditional Phenotypic Value in japonica Rice JIANG Jianhua1,2, CHEN Lan1, LIU Qiangming1, HE Yingjun1, HONG Delin1,* 2012, 26(5): 542-548.  DOI: 10.3969/j.issn.10017216.2012.05.005 Abstract ( )   PDF (400KB) ( )   The elite alleles in carrier varieties for growth duration (GD) and productive panicle number per plant (PN) were detected in order to provide genetic information and breeding materials for breeding  hybrid japonica crosses with wide adaptability and strong competitive advantage of yield. Natural population, composed of 94 japonica varieties, was used as experiment material. Phenotypes of GD, PN and plant height (PH) were investigated in two environments. The conditional phenotypic data were transferred by the linear model method in software QGAStation 1.0, then association analysis based on the unconditional and conditional phenotypes of GD and PN were analyzed by using general linear model in the software TASSEL.A total of 34 SSR marker loci associated with GD and PN were detected in two environments. Among them, 15 were associated with GD, 19 were associated with PN. Four elite alleles of RM8095120 bp, RM7102176 bp, RM72170 bp and RM72178 bp were associated with GD, and their carrier varieties were Hongmangshajing, Nipponbare, Hongmangshajing, Nannongjing 62401, respectively. These elite alleles can shorten GD by 2.03-9.93 d when they were introduced into improved materials. RM72182 bp associated with PN was an elite allele, and its carrier variety was Xiaoqingzhong. RM72182 bp introduction can increase PN by three. What’s more, these elite alleles above can be used to improve target traits without influencing another two traits.

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Molecular  Classification  of   indicajaponica  Rice According to     Widecompatibility Gene S5  Sequence with Endonuclease CEL Ⅰ  NI Shen1 , XIAO Wenfei2, CHEN Hongqi1, WANG Yuexing1, ZHU Xudong1，* 2012, 26(5): 549-554.  DOI: 10.3969/j.issn.10017216.2012.05.006 Abstract ( )   PDF (972KB) ( )   Cloning of the major locus S5，  a widecompatibility gene has laid a molecular basis for identification of the widecompatibility varieties(WCV). According to the difference in S5 locus sequence among indica, japonica and WCV,  PCR was conducted and celery endonuclease CEL Ⅰ,which cleaves DNA with high specificity at sites of basesubstitution mismatch, was used to digest PCR products so as to distinguish various S5 DNA fragments. The results showed that the method is accurate in indicajaponica classification. At the same time, a new genotype on the S5 locus was found.

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On the Mode of Nitrogenreduction in Doublerice Cropping Region in Red Soil Area of South China HOU Hongqian1,2， JI Jianhua1,2， LIU Guangrong1，2， LIU Yiren1， 2， LIU Xiumei1， 2,*， CHENG Zhengxin3， YANG Juncheng4， WEN Shilin5 2012, 26(5): 555-562.  DOI: 10.3969/j.issn.10017216.2012.05.007 Abstract ( )   PDF (466KB) ( )   The grain yield and its components, economic benefits and soil fertility level were compared under eight treatments, namely zero nitrogen (N0), normal nitrogen level(NNL)，80% NNL (chemical fertilizers )and 20% farmyard manure (80F＋20M), 80% NNL (chemical fertilizers), 30% NNL(chemical fertilizers )and 50% farmyard manure (30F＋50M), 30% NNL(chemical fertilizers) and 50% controlledrelease nitrogen (30F＋50S), 30% NNL(chemical fertilizers) and 50% green manure nitrogen in early rice and 80% NNL(chemical fertilizers )and fresh rice straw in late rice, and 80% controlledrelease nitrogen in a 3year location test in South Jiangxi Province of  China with the same  P and K application levels. Under 30F＋50M treatment and 30F＋50S treatment, the grain yield were 4.7% and 5.2% higher than that in 80F, respectively,  equal to NNL treatment. There was obvious difference in yield components factors among different nitrogen treatments. The effective panicle number per hectare  and the spikelet number per panicle were significantly positively correlated with grain yield and seed setting rate was negatively correlatrrd with grain yield.  Soil organic matter, total nitrogen, available nitrogen, available phosphorus contents and pH value in treatment 30F + 50M were higher than that in NNL treatment by 21.3%, 7.0%, 34.7%, 4.9% and 41%. The organic matter, available phosphorus, pH value were significantly higher than that in 80F treatment. There was no significant difference in other nitrogen reduction treatment compared with 80F treatment. Therefore, 30F + 50M, 30F + 50S were recommended as the methods of fertilizer application with nitrogen reduction of 20% in doublerice cropping region in South China.

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Rice Root Growth and Auxin Concentration in Response to Phosphate Deficiency HUANG Rong1， SUN Huwei1， LIU Shangjun1， SONG Wenjing1， LIU Yanxun1， YU Chao1， MAO Ying2, ZHANG Yali1,*， XU Guohua1 2012, 26(5): 563-568.  DOI: 10.3969/j.issn.10017216.2012.05.008 Abstract ( )   PDF (1490KB) ( )   The biomass of rice plant and the initiation and elongation of seminal, adventitious and lateral roots were recorded under five P concentrations (0, 10, 50, 100, 300 μmol/L).  And auxin concentration and relative expression  level of OsPIN genes were also investigated under 10 and 300 μmol/L P concentrations.  Rice shoot biomass decreased with decreasing P concentration, finally resulting in significant increase in root/shoot ratio. Compared with that under 300 μmol/L P concentration, the root/shoot ratio increased by 100% under 10 μmol/L P concentration. The length of seminal, adventitious and lateral roots increased significantly with decreasing P concentrations. However, compared with  that under 300 μmol/L P concentration, the number of adventitious roots and the density of lateral roots decreased with decreasing P concentrations. Compared with that under 300 μmol/L P concentration, auxin concentration increased in the first leaf from top, junction and roots by 85%, 161% and 86% under 10 μmol/L P concentration, respectively. And relative expression  level of OsPIN5a in rice root markedly increased  after exposure to  low P stress for 24 and 96 hours as compared with that under 300 μmol/L P concentration. Enhanced auxin synthesis and polar transport from the shoot to root might be involved in the response of rice to low P stress.

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Osmotic Regulation Substance Contents and Activities of Protective Enzymes in Leaves of Different Hybrid Rice  Combinations as Affected by Shading LIU Li1 , WANG Li1 , DENG Fei1 , HUANG Yun1 , LIU Daiyin2 , REN Wanjun1,* , YANG Wenyu1 2012, 26(5): 569-575.  DOI: 10.3969/j.issn.10017216.2012.05.009 Abstract ( )   PDF (1115KB) ( )   Effects of shading at flowering and grain filling stages on osmotic regulation substance contents and activities of protective enzymes in leaves of different hybrid rice combinations  were investigated by using Ⅱyou 498, Gangyou 188, Dexiang 4103, Gangyou 527 and Chuanxiang 9838 as test materials. The results showed that the content of malondialdehyde of Gangyou 188 was significantly increased under shading, whereas the content of malondialdehyde of Ⅱyou 498, Dexiang 4103, Gangyou 527 and Chuanxiang 9838 had insignificant difference compared to the control. The content of soluble sugar was significantly decreased in Ⅱyou 498, Gangyou 188, Dexiang 4103 and Chuanxiang 9838 under shading. However, the content of souble sugar was not significantly increased in Gangyou 527 under shading. The content of soluble protein in the leaves of different  hybrid rice combinations tended to decrease  under shading. The activities  of protective enzymes in leaves varied among  different  hybrid rice combinations  under shading. Taking both the contents of osmotic regulation substances and the activities  of protective enzymes in rice leaves  into consideration, Gangyou 188 and Gangyou 527 had strong ability to adapt themselves   to weak light conditions. The damage mechanism of leaves of different hybrid rice combinations under shading was discussed.

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Spatial  Distributions of Soil Organic Carbon and Total Nitrogen Concentrations, and Their Influencing Factors in Terraced Paddy Fields of Yuanyang County, China LI Fengbo1, LU Guangde2, ZHOU Xiyue1, YUE Chao2, XU Chunchun1, NI Huixiang2, YANG Xiumei2, FANG Fuping1,* 2012, 26(5): 576-582.  DOI: 10.3969/j.issn.10017216.2012.05.010 Abstract ( )   PDF (806KB) ( )   Spatial distributions of soil organic carbon (SOC) and total nitrogen (TN) concentrations, and their influencing factors in terraced paddy fields of Yuanyang County  were analyzed by geostatistics and  variance analysis.  The SOC and TN  concentrations  in the surface layer(0-20 cm) were 21.38 g/kg and 1.65 g/kg, and moderately variation(36.90% and 32.95%), respectively, and demonstrated moderate spatial dependence according to the nugget ratios (0.67 and 0.71). The semivariogram of SOC and TN were bestfitted by spherical models.   SOC and TN concentrations  increased  with  rising elevations, while decreased with increasing fertilizer inputs.  SOC and TN concentrations   were improved with the excessive fertilization, but  had a negative impact on  the   fertilizer use  efficiency. In all,   attentions should be paid to strengthening  agricultural production management  for improving  land productivity  according to spatial distributions of soil nutrients.

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Impact of Continuous Cross Inoculation on Variation of Xanthomonas oryzae pv.   oryzae from japonica Rice in Yunnan Plateau YANG Yayun1，3,4,#, PENG Xinxi1,2，#, XU Furong1,3,4, HE Jianping1,2, A Xinxiang1,3,4, DONG Chao1,3,4, ZHANG Enlai1,3,4, ZHANG Feifei1，3,4, TANG Cuifeng1,3,4, DAI Luyuan1,3,4,* 2012, 26(5): 583-591.  DOI: 10.3969/j.issn.10017216.2012.05.011 Abstract ( )   PDF (3949KB) ( )   To  study the  factors   influenceing  variation of Xanthomonas oryzae pv. oryzae( Xoo) from japonica rice in Yunnan plateau, eight   strains(30 isolates)   derived from two original strains 20012 and K1  were obtained through successivecross inoculation.Their pathotype and gene variation were studied by using differential cultivars and molecular detection methods. The results showed that  seven obtained strains including 23 isolates observably differed with the  original isolate 20012 in pathotype, beside the pathogenicity of 62 containing 3 isolates were increased, while the pathogenicity of  the other six strains containing 20 isolates were decreased. The pathotypes of 68 were the  same as that of its original isolate K1. By using UPGMA analysis of PCR producets of primer  IS1112  and  hrpF, no   relationship between molecular haplotypes and pathotypes were found. Sequence analysis showed that basic group variation  in sequences of all strains amplified by primers  IS1112  and  hrpF differed. The pathogenic variation of obtained strains from two original strains  inoculated on various varieties were unique and nondirective.

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Identification of Pita and Pib Genes for Rice Blast Resistance of Rice Landraces from Yunnan Province LI Jinbin1, WANG Tian2, XU Minghui2,* 2012, 26(5): 592-598.  DOI: 10.3969/j.issn.10017216.2012.05.012 Abstract ( )   PDF (514KB) ( )   The existence of the two genes of Pita and Pib in 176 rice landraces from Yunnan Province were rapidly determined using the dominant DNA markers of Pita/pita and Pib/pib alleles(YL155/YL18 and YL183/YL87 for the former, PibdomF/Pib dom R and Lys14SF/Lys145R for the latter designed according to their difference in sequence). The results showed that Pita  was found in 72 among 176 landraces with the frequency of  40.9%. Among them,  Pita  was found in 44 among 77 indica and 28 among 99 japonica cultivars, the frequency in indica and japonica were 57.1% and 28.3%, respectively. The distribution frequency of the resistance gene Pita in the japonica and indica varieties significantly differed, the frequency was higher in indica than in japonica cultivars. While Pib was found in 33 among 176 landraces with the frequency of  18.8%. Among them, Pib was found in 15 indica and 18 japonica cultivars, the frequency in indica and japonica were 19.5% and 18.2%, respectively.  Pita was mainly distributed in Yuxi, Baoshan, Lincang and Simao prefectures, while Pib was mainly distributed in Nujiang, Baoshan and Zhaotong prefectures of Yunnan Province. The results of pathogenic assays with the rice blast fungus in greenhouse were generally consistent with the results of molecular identification.

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Component Analysis, Activity Measurement and Pathogenic Effect of Cell Wall Degrading Enzymes from Rice Sheath Blight Pathogen Rhizoctonia solani YANG Mei1，#, YANG Yingqing1,2，#, ZHENG Li1, LI Minghai1, ZHOU Erxun1,* 2012, 26(5): 599-605.  DOI: 10.3969/j.issn.10017216.2012.05.013 Abstract ( )   PDF (1069KB) ( )   The kinds of cell wall degrading enzymes (CWDEs) from Rhizoctonia solani and their pathogenic effect were analyzed by the SDSpolyacrylamide gel electrophoresis (SDSPAGE), enzyme activity measurement, determination of leaky reducing sugar and relative conductivity of leachate from leaf tissues treated with CWDEs. The results of SDSPAGE of CWDEs revealed that there were at least 10 clear bands of proteins, indicating that there were at least 10 kinds of components with different molecular weight in the mixture of CWDEs. The enzyme activities of seven kinds of common CWDEs’ were determined, and the results showed that the activity of polygalacturonase (PG) was the highest, followed by polymethylgalacturonase (PMG). Through comparing the effects of different substrates on the induction of three enzymes, it was found that 1.0% sodium carboxymethyl cellulose (CMC) was the most suitable substrate for the induction of endoβ1,4glucanase (Cx), whereas 1.0% pectin for the induction of PG and PMG. The best enzyme reaction conditions for the three most important CWDEs, i.e. PG, Cx and PMG, were explored from three aspects including temperature, reaction time and pH value. In regarding to reaction temperature, it was found that the best reaction temperature was 30℃ for Cx,  and  50℃ for PG and PMG. In regarding to reaction time, it was found that 40 min was more suitable for Cx and PMG, whereas 60 min was more suitable for PG. In regarding to pH value, the results indicated that the enzyme activities of PG, Cx and PMG were the highest when the pH values reached 5.0, 9.0 and 4.0 to 6.0, respectively. Through the determination of leaky reducing sugar and relative conductivity of leachate from leaf tissues treated with CWDEs, it was obvious that CWDEs did cause damage to rice leaf tissues and the damage degrees increased with the increasing concentrations of CWDEs.

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Research Trends of Rice Science Based on Web of Science SUN Xiuhuan， LU Wenru* 2012, 26(5): 606-614.  DOI: 10.3969/j.issn.10017216.2012.05.014 Abstract ( )   PDF (454KB) ( )   Worldwide published rice articles during 2001 and 2010 were analyzed based on the Web of Science database by the bibliometric method. Especial concerns were paid on the highly influential countries/regions, research institutes, journals, authors and papers; and the rice research hotspots were summed up based on the ESI Web of Science database. The USA, Japan, China were leading the rice research. Institutions from Japan and China issued the largest amount of rice papers, while the  papers  from the USA were higher qualified and more influential. The core journals that carried rice papers were mainly in the USA, England, the Netherlands and Germany. Nine of the 20 most frequently cited papers were from the USA, and 3 from Japan and China, respectively. The research hotspots in rice were transgenic rice, QTL, rice genome, rice blast, cultivated rice, reactive oxygen species, arsenic, iron, drought resistance    and so on.

Experimental Techniques

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Reference Gene Selection of Ustilaginoidea virens by Realtime  PCR GU Zhimin1,2, DING Zhengzhong1， CHEN Xifeng1， GUO Longbiao2， ZENG Dali2， QIAN Qian2,*， MA Bojun1,* 2012, 26(5): 615-618.  DOI: 10.3969/j.issn.10017216.2012.05.015 Abstract ( )   PDF (1793KB) ( )   A total of five traditional reference genes of Ustilaginoidea virens were systematically compared by using realtime quantitative PCR, including 18S rRNA, GAPDH, ubiquitin, βactin and αtubulin. The expression stability of the 5 candidate reference genes was evaluated with geNorm program when the fungus was 7, 8, 9 and 10 days old or exposed to 0.4 mol/L NaCl for 0 h，4 h， 8 h， 12 h， 16 h. The results showed that αtubulin2 and βactin were stably expressed when the fungus was at different days of age. For the case of 0.4 mol/L NaCl stress, βactin and αtubulin2 were also expressed stably. As a conclusion, βactin and αtubulin could be used as the reference genes for normalization of realtime quantitative PCR data.

Short Communications

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Identification of Multiobjective Rice Lighttrap Pests Based on Template Matching LV Jun1 , YAO Qing1, LIU Qingjie1, XUE Jie1, CHEN Hongming3, YANG Baojun2, TANG Jian2,* 2012, 26(5): 619-623.  DOI: 10.3969/j.issn.10017216.2012.05.016 Abstract ( )   PDF (899KB) ( )   Identification and count of rice lighttrap pests are very important in monitoring rice pests. The pests trapped by black light lamps show different postures and incomplete bodies, which increase the difficulty of image automatic identification.  Template matching and Kfold cross validation methods were used to identify multiobjective rice lighttrap pests based on nontouching pest images. Firstly, one hundred and fiftysix features including color, shape and texture features were extracted from each pest image. Secondly, the principal component analysis was employed for reducing data dimensionality, and first six principal components were selected as pests’ features. Then, the template number was determined according to the gesture of each pest species  and the template parameters were obtained from the cluster centers by fuzzy Cmean clustering method. Finally, single and multitemplate matching methods were used to identify rice pests. The results showed that the accurate rate of multitemplate matching and single template matching were 83.1% and 59.9%, respectively for rice lighttrap pests with multiple postures and some incomplete bodies.

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AMMI Model Based Analysis on Head Milled Rice Rate of Hybrid indica Rice in China LU Lin， ZHU Zhiwei*， DUAN Binwu 2012, 26(5): 624-628.  DOI: 10.3969/j.issn.10017216.2012.05.017 Abstract ( )   PDF (499KB) ( )   Head milled rice rate (HMRR) of six series of indica hybrid rice from five rice growing regions was analyzed by using additive maineffects and multiplicative interaction(AMMI) model. The stability and adaptability of HMRR of  indica hybrid rice were evaluated by analysis on genotype, environment effect and genotype×environment interaction. The results showed that Liangyoupeijiu had higher HMRR and excellent stability. HMRR of Ⅱyou 838 and Fengliangyou 1 was higher but less stable. Dyou 527 and Jinyou 207 had low HMRR but better stability. HMRR of Gangyou 725 was low and worst stable. Dyou 527, Gangyou 725 and Jinyou 207 performed better adaptability in rice growing region of Zhejiang,Shanghai and Jiangsu. Liangyoupeijiu, Ⅱyou 838 and Fengliangyou 1 showed good adaptability in other four  regions. The rice growing region of Anhui,Jiangxi,Hubei and Hunan and rice growing region of Yunnan,Guizhou,Sichuan and Chongqing showed weak interaction and ability to distinguish cultivars. It is concluded that genotype×environment interaction effect has greater influence on HMRR.

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Expression Characteristics of the Ospgip1 Gene in  Rice Cultivars  with Resistance or Susceptiblility  to Sheath Blight CHEN Xijun1, 2, LIU Xiaowei1, ZUO Shimin2, TONG Yunhui1, PAN Xuebiao2,*, XU Jingyou1，* 2012, 26(5): 629-632.  DOI: 10.3969/j.issn.10017216.2012.05.018 Abstract ( )   PDF (569KB) ( )   A  fragment of 1 184 bp of the Ospgip1 gene including a complete  open reading frame (OFR) of 930 bp  had been amplified, which had the termination codon of TAA and no intron.  RTPCR results showed that the Ospgip1 gene could be expressed in all the rice cultivars while their expression levels differed among growth stages or organs. Realtime PCR results indicated that the expression levels in resistant cultivars YSBR1 and Jasmine 85 were  significantly higher than  that in the susceptible cultivar Lemont. The expression level was also upregulated in the etiolated seedlings. Inoculation with the pathogen of rice sheath blight significantly upregulated the expression level of the  Ospgip1 gene in resistant cultivars  while  exerted no  impact on the expression level of the gene in  the susceptible one.

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Safety Assessment of Bt63, a Genetically Modified Rice in vitro CHEN Wujian1, ZHENG Wei2,*, LU Qinfeng2, ZHANG Mingzhe1, WU Zhonghua2, XU Qi2, WU Zhiyi1, LI He2 2012, 26(5): 633-636.  DOI: 10.3969/j.issn.10017216.2012.05.019 Abstract ( )   PDF (336KB) ( )   To investigate the cytogenotoxicity of Bt63, a genetically modified(GM) rice in human lymphocytes  in vitro,  human lymphocytes were exposed to whole protein of Bt63 at doses of 200, 100, 50, 25, 12.5 μg/mL and incubated for 1.5 h, 6 h and 24 h, respectively. The DNA damage, chromosomal aberrations and cytotoxicity induced by whole protein of Bt63 were measured by comet assay, micronucleus test, CCK8 assay and neutral red uptake (NRU) assay. After different incubation periods, the cytogenotoxicity of lymphocytes in positive control group was significant higher than that of lymphocytes in blank control group. Moreover, the exposure timeeffect relationship was observed in positive control group by CCK8 assay and neutral red uptake (NRU) assay. There was no significant difference in cytogenotoxicity between Bt63 group and nonGM rice group. Also, the Bt63 GM rice group did not show higher cytogenotoxicity than that of nonGM rice group, indicating that Bt63 GM rice has similar cytogenotoxicity with nonGM rice, and Bt63 GM rice has no cytogenotoxicity effect on human lymphocytes in vitro.  Compared with traditional toxicological  methods, the experimental system is economic, simple, rapid and reliable.