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10 November 2014, Volume 28 Issue 6 Previous Issue    Next Issue
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A Proteomic Study on a DiseaseResistanceEnhanced Rice Lesion Mimic Mutant HAN Xueying1,2, YANG Yong2, YU Chulang2, ZHANG Wenhao3 , YE Shenghai2, CHEN Bin2, CHENG Chen2,CHENG Ye2, YAN Chengqi2, CHEN Jianping1,2,* 2014, 28(6): 559-569.  DOI: 10.3969/j.issn.1001-7216.2014.06. 001 Abstract ( )   PDF (2007KB) ( )   A lesionmimic mutant in rice (Oryza sativa L.), chloroplasticH2O2induced lesion 1 (chl1), has enhanced resistance to rice blast and bacterial blight. To understand the molecular mechanisms underlying this phenotype and its resistance responses, a proteomicsbased approach was used to identify  differentiallyexpressed proteins between chl1 and its wild type. Using twodimensional fluorescence difference gel electrophoresis technology and mass spectrometry, 70 protein spots were successfully identified, of which 46 were upregulated and 24 were downregulated in the mutant. These differentiallyexpressed proteins are involved in diverse biological processes including disease resistance, photosynthesis, oxidationreduction reaction, amino acid/protein metabolism, chaperoning and carbohydrate metabolism. The complex regulatory network in which these proteins are involved may play an important role in regulating the programmed cell death and the resistance reaction in chl1.

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QTL Mapping of Heading Date and Yieldrelated Traits in Rice Using a Recombination Inbred Lines (RILs) Population Derived from BG1/XLJ ZHAN Xiaodeng， YU Ping， LIN Zechuan， CHEN Daibo， SHEN Xihong， ZHANG Yingxin， FU Junlin， CHENG Shihua*， CAO Liyong* 2014, 28(6): 570-580.  DOI: 10.3969/j.issn.1001-7216.2014.06.002 Abstract ( )   PDF (2121KB) ( )   Heading date and yieldrelated traits including plant height (PH), number of tillers per plant (NTP), panicle length (PL), number of filled grains per panicle (NFGP), grain number per panicle (GNPP) and seed setting rate (SR) are important agronomic traits in rice. Detection of new QTL associated with heading date and yieldrelated traits have great significance in new genes identification and utilization of rice. We used a recombination inbred lines (RILs) population derived from BG1/XLJ to detect QTL associated with heading date and yieldrelated traits in two environments, effects of epistatic and interaction with environments were also analyzed. A total of 32 QTL associated with heading date and yield traits were detected on chromosomes 2, 3, 4, 5, 6, 7 and 8. Furthermore, five pairs of QTL intervals had significant twofactor epistatic effects, and 5 QTL had significant interaction with environments. Many yield related QTL detected were identified near the intervals where yieldrelated QTL were detected in previous researches. Among them, the QTL with large effects commonly clustered. Low epistatic and interaction with environments effects for yield related QTL, indicated small interactive effects between genes, genes and environments.

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Sequence Analysis and Effect of Grain Width Genes GW2 in Rice Based on Extreme Materials in Grain Size ZHANG Yadong1,2, LIANG Yanli1,2, ZHENG Jia2, DING Dan1,2, ZHAO Chunfang2, CHEN Tao2, ZHAO Qingyong2, ZHU Zhen2, ZHOU Lihui2, YAO Shu2, ZHAO Ling2, YU Xing2, WANG Cailin2,* 2014, 28(6): 581-588.  DOI: 10.3969/j.issn.1001-7216.2014.06.003 Abstract ( )   PDF (4464KB) ( )   Grain shape is one of the most important influencing factor to rice grain yield. Among all of the cloned grain shape genes, GW2 is an important one controlling grain width. We obtained the sequences of the important grain shape gene GW2 using cDNA cloning strategy from rice variety TD70 and Kasalath and design its functional marker based on gene sequence difference to analyze genotype and phenotype of recombinant inbred lines(RILs) from TD70/Kasalath cross, some indica restorer lines and japonica rice cultivars in Jiangsu Province.  GW2TD70 shared 100% identity to GW2WY3 and 99.77% identity to GW2 sequence（GenBank: EF447275）and there exist a single nucleotide mutation at site 114 without changing amino acid, a single nucleotide deletion at site 316 which generate termination codon at site 346, 347 and 348,thus the expression of GW2 protein stopped in advance. The sequence of GW2Kasalath showed 99.92% identity to GW2 which is known, and there exists a single nucleotide change at site 114 without the variation in amino acid. Comparing with GW2 in TD70, the gene in Kasalath express the intact protein without advanced stop. For GW2 gene, the dCAPS marker was designed according to a base deletion of A in the 316 site of sequence. Digested by Apo I, TD70 showed 21 bp and 30 bp fragments and Kasalath showed 51 bp fragment. Among the 240 RILs, 82 contained GW2TD70 and 158 contain GW2Kasalath , there are significant differences between their phenotypic value. No GW2TD70 gene exit in 10 japonica rice cultivars from Jiangsu Province and 8 indica restorer lines. GW2 gene derived from TD70 may have regulatory effects on grain width，and we can use it to improve grain weight in rice breeding.

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QTL Analysis for Rice Leaf Morphology at Maturity Stage Using a Recombinant Inbred Line Population Derived from a Cross Between Nipponbare and 9311 WANG Lan, HUANG Lichao, DAI Liping, YANG Yaolong, XU Jie, LENG Yujia, ZHANG Guangheng, HU Jiang, ZHU Li, GAO Zhenyu, DONG Guojun, GUO Longbiao, QIAN Qian, ZENG Dali* 2014, 28(6): 589-597.  DOI: 10.3969/j.issn.1001-7216.2014.06.004 Abstract ( )   PDF (2490KB) ( )   As the main photosynthetic organs, rice leaf morphological characteristic is an important target in high yield breeding. A recombinant inbred line (RIL) population derived from a cross between Nipponbare and 9311 was used to map QTLs  for leaf length, leaf width, leaf area and leaf length to width ratio of top three leaves in maturity stage. Correlation analysis showed that there were significant correlation among leaf shape indexes. The leaf shape traits followed a normal distribution with some transgressive individuals,indicating that these complex quantitative traits were controlled by multiple genes. A total of 42 QTLs were detected for these traits, which were mapped on all 12 chromosomes with the phenotypic variations ranging from 4.3% to 29.0%, and the increasing effective alleles contributed by 9311 showed higher additive effect than that from Nipponbare. Seventeen pairs of QTL with epistatic effect were identified among eight traits with explained variations from 3.3% to 23.4%. Pyramiding the positive alleles apparently altered the characters of flag leaf.

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QTL Analysis on New Root Traits after Rice Transplanting JIANG Shukun, ZHANG Fengming*, BAI Liangming, SUN Shichen, WANG Tongtong, DING Guohua, JIANG Hui, ZHANG Xijuan 2014, 28(6): 598-604.  DOI: 10.3969/j.issn.1001-7216.2014.06.005 Abstract ( )   PDF (684KB) ( )   To reveal the genetic basis of the new root  traits after transplanting, by using Sasanishiki, Habataki and 85 backcross recombinant inbred lines (BC1F5) and its linkage map including 245 molecular markers QTL controlling new root   traits after transplanting were detected. Ten QTLs controlling average diameter, total root length, average surface area, root length and root number were identified. These QTLs distributed on chromosome 1, 2, 5, 9 and 11. The phenotypic variations (VE) explained by individual QTL ranged from 10.7% to 28.5%. QTL clusters were formed on chromosome 9 and 11. Then four major QTLs were fine mapped by “selective mapping” method. The QTLs (qRL11.1，qASA11 and qRN11.1) for root length, average surface area and root number were located between C477 and G320B (5 461 121-6 686 166 bp) on chromosome 11. It was a novel locus with research and application value.

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Delayed Inheritance of Purple Pericarp in Rice   and Development of Functional Marker for Pb Gene WANG Fangquan1,2, YANG Jie1,*, FAN Fangjun1, WANG Jun1, ZHU Jinyan1, LI Wenqi1, SHEN Wenbiao2,ZHONG Weigong1 2014, 28(6): 605-611.  DOI: 10.3969/j.issn.1001-7216.2014.06.006 Abstract ( )   PDF (1754KB) ( )   In order to determine the heredity of purple pericarp, the purple pericarp cultivar Xiangxuenuo, developed in Jiangsu Province, and white pericarp cultivar 02428 were used as parents to make reciprocal crosses, and the colour of the seeds of relevant F1, F2, F3 generation were phenotyped. The results indicated that the purple pericarp of Xiangxuenuo was controlled by single dominant nuclear gene, belonging to the typical maternal influence. Purple pericarp in rice was due to a 2 bp deletion in Pb gene on chromosome 4 which encoded a basic helixloophelix protein. A CAPS marker was developed based on the 2 bp deletion, and the CAPS  marker was cosegregated with the colour of pericarp in the F2 population, allowing to identify the homozygous purple pericarp genotype, homozygous white pericarp genotype and heterozygous purple pericarp genotype individuals. The PCR products of 10 rice lines were sequenced. All the purple pericarp rice lines carried GT deletion. It was interesting that the wild rice lines with red pericarp carried the normal allele; therefore we speculated that the purple pericarp was mutated from cultivated rice. Furthermore, one line with purple hull and white pericarp carried pb allele, it showed the complex genetic mechanism in rice pigmentation. In conclusion, the CAPS  marker is a perfect marker for purple pericarp rice breeding and the evolution of Pb gene need further research.

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Physical and Chemical Characteristics and Lodging Resistance of Culm of indica Hybrid Rice F you 498 under Mechanical  Planting LEI Xiaolong 1, LIU Li1, LIU Bo1, HUANG Guangzhong2, MA Rongchao1,*, REN Wanjun1,* 2014, 28(6): 612-620.  DOI: 10.3969/j.issn.1001-7216.2014.06.007 Abstract ( )   PDF (483KB) ( )   In the twentyfifth day after full heading, the physical and chemical characteristics and lodging resistance of basal internodes in main stem and tillers  were studied under three planting methods at two sowing dates in a split plot field experiment with F you 498 as material. The results showed that lodging resistance of artificallytransplanted rice was significantly higher than that under mechanized planting, and the yield of mechanical  directseeding rice was significantly lower than that of the mechanical  transplanted and the artificial transplanted rice. Correlation analysis indicated that lodging index was positively correlated with gravity center height, internode length, bending moment and contents of N and P, and negatively correlated with internode diameter and thickness, culm type index, dry weight to length ratio of the of unit internode, contents of cellulose, lignin, soluble sugars and K, spikelet number per panicle and grain yield. Compared with artificial transplanting rice,mechanical directseeding and transplanting rice showed higher gravity center height, longer basal internodes and higher contents of N and P, smaller culm type index and RWLI(dry weight to length ratio of the internode). The lodging indexes were larger under mechanized directseeding and transplanting due to its larger bending moment and lower breaking moment. Gravity center height and internonde length increased and the internode diameter and thickness, plumpness status, contents of cellulose, lignin, soluble sugars and K and breaking moment lowered with delaying sowing date. Thus, lodging indexes under late sowing increased significantly. The short, thick and sturdy basal internode and higher contents of cellulose, lignin and K could help strength lodging resistance. The changes in structure and chemical contents of culm due to the changes in planting methods and sowing dates were probably the main reasons for the variation of lodging indexes.

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Difference in Yield and Its Components Characteristics of Different Type Rice Cultivars in the Lower Reaches of the Yangtze River JIANG Yuanhua, ZHANG Hongcheng*， ZHAO Ke, XU Junwei, WEI Huanhe, LONG Houyuan, WANG Wenting, DAI Qigeng, HUO Zhongyang, XU Ke, WEI Haiyan, GUO Baowei 2014, 28(6): 621-631.  DOI: 10.3969/j.issn.1001-7216.2014.06.008 Abstract ( )   PDF (533KB) ( )   With  20 representative rice cultivars belonging to  four types as materials, a field experiment was conducted to reveal the difference in yield and its components in the ricewheat cropping areas(Yangzhou, Changshu) in the lower reaches of the Yangtze River in 2012-2013. Grain yield and its components, stems and tillers and panicle traits and grainfilling were analyzed systematically.Result show that, 1)under the high yielding cultivation conditions, rice grain yield followed a decline trend of indicajaponica hybrid rice(IJHR) >japonica hybrid rice(JHR)>traditional japonica rice(TJR)>indica hybrid rice(IHR)  in different sites and years.The average yield of IJHR in both years in Yangzhou was 12499.4 kg·hm-2, 4.82%,11.94% and 19.68% higher than those of JHR,TJR and IHR.2)The contribution of yield components to yield was total spikelet number>seedsetting rate>1000grain weight,and the contribution to total spikelet number was grain number per panicle>number of effective panicles.It illustrated that big panicle remained the main way for high yield of rice.3)The number of stems and tillers at jointing stage was TJR>IHR>JHR>IJHR,the contribution rates of stems and primary tillers were IJHR>IHR>TJR>JHR, while  the contribution rate of the secondary tillers was JHR>TJR>IHR>IJHR, the composition of stems and tillers at waxy stage was the same as that at jointing stage;percentage of effective tillers of  TJR(7576%) ranked first，followed by JHR(7287%),IJHR(6680%) and IHR(63.24%).4)Panicle length followed a trend of IHR>IJHR>JHR>TJR;grain density,single panicle weight were IJHR>JHR>IHR>TJR;grain number on the primary rachis branch was JHR>IJHR>TJR>IHR,while number of branches and total grain number on the primary rachis branch were IJHR>JHR>TJR>IHR,the trend of the contribution rate of grain number on the primary rachis branch to grain number per panicle from high to low was TJR（41.00%）,JHR（35.50%）,IJHR（31.83%）,IHR（27.92%）;grain number on the secondary rachis branch was IHR>IJHR>TJR>JHR,while number of branches and total grain number on the secondary rachis branch were IJHR>JHR>IHR>TJR,the trend of the contribution rate of grain number on the secondary rachis branch to grain number  per panicle from high to low was IHR(72.80%),IJHR(68.17%),JHR(64.50%),TJR(59.00%). 5)The final grain weight was TJR>JHR>IHR>IJHR，maximum grainfilling rate was IHR>TJR>JHR>IJHR,the time reaching the maximum grainfilling rate was JHR>TJR>IJHR>IHR,mean grainfilling rate was IHR>TJR>JHR>IJHR,effective grainfilling time was JHR>IJHR>TJR>IHR;For grainfilling characteristics during different stages，grainfilling amount of gradual growth stage, fast growth stage and slow growth stage were TJR>JHR>IHR>IJHR;grainfilling time of gradual growth stage was JHR>TJR>IJHR>IHR,grainfilling time of fast growth stage and slow growth stage were JHR>IJHR>TJR>IHR; mean grainfilling rate of gradual growth stage was IHR>TJR>IJHR>JHR,grainfilling rate of  fast growth stage and  slow growth stage were IHR>TJR>JHR>IJHR.Compared with JHR ,TJR and IHR, IJHR had obvious advantages in productivity with big panicle as the basis.

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Effect of Reduced Phosphorus Fertilizer Application on Yield Formation and Quality of japonica Rice in Jiangsu Coastal Region HUANG Lifen, TAO Xiaoting, GAO Wei, WANG Yuanling, ZHUANG Hengyang* 2014, 28(6): 632-638.  DOI: 10.3969/j.issn.1001-7216.2014.06.009 Abstract ( )   PDF (432KB) ( )   Soil available phosphorus content of farmland has reached a high level in Jiangsu coastal area due to longterm application of large amounts of phosphate fertilizer. But current  higher rates of phosphate fertilizer decreases the phosphorus effect, even having  an adverse effect on the surrounding water environment. A field experiment was conducted to investigate the effect of reduced phosphorus fertilizer application on the yield formation and quality in rice. Six treatments were designed as CK(no fertilization), conventional  NPK fertilizer application, no phosphorus fertilization, reduced P rate( 50% of conventional phosphorus fertilization rate), conventional phosphorus fertilization, increased P rate (150% of  conventional phosphorus fertilization). The results showed that there were no significant difference in SPAD of rice leaves among all the treatments except CK at jointing, heading, middle and late filling stages. The dry matter accumulation at jointing, heading stage, maturity stage and net dry matter accumulation as well as net population growth rate during jointingheading, headingmaturing had also no significant difference among all the treatments except CK. The 1000grain weight under no or reduced  phosphorus treatments was significantly lower than that of conventional phosphorus treatments, while the  rest of yield components as well as economic coefficient had no significant difference among the treatments. Due to higher effective panicle number, grain number per panicle and 1000grain weight, the rice grain yield under  reduced phosphorus treatment reached 9 801 kg/hm2, 7.69% higher than that under the conventional phosphorus treatment. There were no significant differences in the indexes of rice quality except that the chalky grain rate under the reduced phosphorus treatment was significantly higher than CK. Due to higher peak viscosity and breakdown and lower setback value, the eating quality of rice under the reduced phosphorus treatment was improved.

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Component Analysis on Volatile from Conventional and Hybrid Rice Leaves During Different Developmental Stages HUANG Min1, HU Fei1,*, YANG Jun2, JI Jinghua1, CHEN Xionghui1 2014, 28(6): 639-648.  DOI: 10.3969/j.issn.1001-7216.2014.06.010 Abstract ( )   PDF (710KB) ( )   Volatile components of conventional and hybrid rice leaves were analysed by solid phase microextraction (SPME) and gas chromatographymass spectrometry(GCMS) with two conventional rice varieties(Allelopathic rice 3, Xiangyaxiangzhan) and two hybrid  rice combinations (Ruanhuayou 3, Nongzhenyou 3) as materials during four developmental stages. The results showed that 41 and 37 components were identified in Allelopathic rice 3 and Xiangyaxiangzhan; 49 and 53 in Ruanhuayou 3 and Nongzhenyou 3. For Allelopathic rice 3, Xiangyaxiangzhan and Ruanhuayou 3, the number of volatile components maximized during milkripe stage, while for Nongzhenyou 3 during fullripe stage. There were some insignificant differences in volatile components and their relative content between conventional rice and hybrid rice combination during the same stage.  The relative content of alkanes and terpenes decreased with rice ripening in the four tested rice. Longifolene and Squalene were identified in all tested rice during different stages. (Z)3Hexenol from Ruanhuayou 3 in flowering stage and γCaprolactone from Xiangyaxiangzhan and Nongzhenyou 3 in fullripe stage were identified, these two components were green leaf volatiles.The relationship between these components and rice quality, pests was worth of further study.

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Genomewide Association Analysis on Genes Controlling Panicle Traits of Varieties from International Rice Core Collection Bank and Its Breeding Utilization ZUO Shimin1,2, KANG Houxiang2, LI Qianqian1, CHEN Zongxiang1, ZHANG Yafang1, LIU Wende2, WANG Guoliang2, CHEN Hongqi3, *, PAN Xuebiao1,* 2014, 28(6): 649-658.  DOI: 10.3969/j.issn.1001-7216.2014.06.011 Abstract ( )   PDF (2598KB) ( )   Genomewide association study (GWAS) on five panicle traits of 315 rice accessions introduced from international rice microcore germplasm bank was conducted. The  panicle length (PL) and the primary rachis  branch number (PBN) exhibited significant difference between the two plant locations, while  grain length (GL), grain width (GW) and grain length to width ratio (GLWR) were not, indicating that PL and PBN are sensitive to environments. Totally 7, 5, 10, 8 and 6 chromosome regions or SNP (single nucleotide polymorphism) marker loci were significantly associated with PL, PBN, GL, GW and GLWR, respectively. Among them, eleven regions associated with grain shape and one region controlling PBN  were  consistence in two environments. Significant linear correlation was found between the average trait value and the number of favorable alleles carried by the cultivars in all associational loci. Among all associational loci, the cultivars in Aromatic (ARO) and tropical japonica (TRJ) subpopulations owned most favorable alleles, but those in temperate japonica (TEJ) subpopulation contained the least. The domestic varieties mainly harbored unfavorable alleles in 6 of associational loci detected. Totally 15 varieties from 11 different countries harbored more than 30 favorable alleles, which were fallen in TRJ subpopulation, demonstrating  that the cultivars in TRJ subpopulation have high potential in rice breeding for high and stable yield. Based on these results, we discussed the approach to improve domestic rice panicle traits by using TRJ varieties.

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Research on Magnaporthe oryzae Induced Alternations of Ion Absorption in Roots of Susceptible and Resistant Rice Varieties HOU Peichen1, WANG Cheng1,* , WANG Xiaodong1, WANG Meijuan2, ZHU Dazhou1, GAO Quan1 2014, 28(6): 659-664.  DOI: 10.3969/j.issn.1001-7216.2014.06.012 Abstract ( )   PDF (1156KB) ( )   The dynamic ion flux test technique was employed to analyze ion absorption by roots in susceptible and resistant rice varieties subjected to Magnaporthe oryzae infection,aiming to determine the effect of Magnaporthe oryzae infection on nutrient absorption in rice plants. The results showed that Magnaporthe oryzae infection caused more obvious disease symptom in susceptible variety and reduced K+ absorption, led to H+ transition from efflux to influx, and reduced Ca2+ absorption with larger unstability  in root. In contrast, the resistant rice variety showed moderate disease severity and was characterized by only a slight decrease in absorption of K+, H+, and Ca2+ ions by the roots. These results indicated that subjected to Magnaporthe oryzae infection, resistant rice variety showed stronger ion absorption capacity, while the susceptible rice variety exhibited weaker capacity, which could be an important reason leading to the barrier of nutrients absorption in the roots of susceptible rice variety.

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Laboratory  Identification of Resistance to Pesticides and rpfC Gene Sequence Analysis of Xanthomonas oryzae pv. oryzae  in japonica Rice from Yunnan Plateau YANG Yayun1, ZHANG Enlai1, A Xinxiang1, TANG Cuifeng1, ZHANG Feifei1, CHEN Dan1, DONG Chao1,WU Kun1,2, XU Furong1, ZHOU Ying3, QI Chunxue4, DAI Luyuan1,* 2014, 28(6): 665-674.  DOI: 10.3969/j.issn.1001-7216.2014.06.013 Abstract ( )   PDF (3246KB) ( )   In order to reveal pesticide resistance and its mechanisms of Xoo strains belonging to 10 pathogenic types in japonica rice from Yunnan plateau, the indoor resistance identification of Xoo strains to pesticides were conducted in NA medium containing pesticides at different concentrations, PCR amplification was carried out using specific rpfC gene primers which was  closely related to resistance, gene and amino acid were sequenced and aligned. The results show that the minimun inhibitory concentration(MIC) of bismerthiazol  to all tested strains were 40180mg/L, while the  MIC  of  Benzoic acid1, 3dithiolan2ylidenehydrazide(BADYH, IV)  to the strains were 10100mg/L, there was no resistant strains to stretomycin+oxyteracykine. Correlated with the sensitivity to pesticide, the pathogenicity of Xoo strains was improved with the rising resistance to pesticide. The homology alignment results showed that rpfC gene of 10 strains shared 92%-98% identity with the gene of the registration number X97865.1 in the Genebank, while the differences in 11 RpfC protein sequences ranged from 8.3% to 99%. There were six complete functional domains in RpfC protein of strain 200131 with the strongest pathogenicity（Type Ⅵ）and resistance to pesticide. REC reception domain in RpfC protein of strain DHL1 has disappeared , which showed the weakest pathogenicity（Type 0）and resistance to pesticide. Particularly, strains ⑤ (Type Ⅳ), middle pathogenic and resistant,could not form functional domains of RpfC protein.

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Identification and Analysis on the Rice Blast Resistance Gene Pita in Wild Rice from Yunnan， China ZHAO Guozhen1, JIANG Chunmiao2, LIU Jixin1 , CHEN Yumin1, YU Tengqiong2, CHENG Zaiquan2,* 2014, 28(6): 675-680.  DOI: 10.3969/j.issn.1001-7216.2014.06.014 Abstract ( )   PDF (742KB) ( )   A total of 16 accessions of wild rice from different regions of Yunnan Province were used for PCR amplification with Pita gene specific primers. The amplified PCR products were cloned into Topo cloning vector, and sequenced.  Among 16 accessions, the identical sequence fragment to the resitance Pita gene was amplified separately from 7 accessions. The Pita allele in one accession of Oryza rufipogon with purple stem from Jinghong contains an identical DNA sequence in the coding region to the original Pita resistance gene. It suggested that Yunnan might be one of the origins of the Pita gene.