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Reverse Genetics Analysis of the Transcription Factor OsHox9, a Member of Homeobox Family, in Rice AI Liping, SHEN Ao, GAO Zhichao, LI Zhenglong, SUN Qionglin, LUAN Weijiang* 2014, 28(3): 223-228.  DOI: 10.3969/j.issn.1001-7216.2014.03.001 Abstract ( )   PDF (1114KB) ( )   The homeobox transcription factors play important roles in the growth and development of the plants, involved in regulation of cell differentiation, the architecture of morphogenesis and response to environmental signals in plants. To study the function of these transcription factors in rice, we constructed RNAi vectors of OsHox9 and analyzed the function of OsHox9 by the reverse genetics method. The results showed that the height and tiller number of RNAi transgenic plants were decreased in comparison with wild type plants. Realtime PCR analysis showed that OsHox9 expression level was  downregulated in transgenic plants with phenotype. However, the expression level of OsHox9 in transgenic plants without phenotype was similar to wild type plants, suggesting that the phenotypes of transgenic plants were caused by RNAi effects. In addition, we also analyzed the expression pattern of OsHox9 in different organs of rice. The results showed that OsHox9 was expressed in different organs, especially displayed high expression level  in stem apical meristems and young panicles. To investigate the subcellular localization of  OsHox9, we constructed the fused vector 35S::OsHox9GFP to introduce into leaves of tobacco. The transient expression result showed  OsHox9  was  localized  on the cell membrane.

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Genetic Analysis, Gene Mapping and Breeding Application of Small Grain and Dense Panicle Mutant sep1 in Rice WANG Yuexing1,＃, WU Kun2,＃, FANG Yunxia1, CHEN Hongqi1, NI Shen1, FU Xiangdong2, ZHU Xudong1,* 2014, 28(3): 229-234.  DOI: 10.3969/j.issn.1001-7216.2014.03. 002 Abstract ( )   PDF (1660KB) ( )   By 60Co rray radiation, a small grain and dense panicle mutant sep1 was obtained  from the indica variety Shuangkezao. Compared to the wild type Shuangkezao, it was a mutant with smaller grain, shorter spikestalk but more grain number per panicle.  The histological slice observation of the main stem showed that the mutant had more vascular bundle number in one cycle and more cell number per unit area than the wild type. Genetic and allelism analysis results showed that the small grain and dense panicle mutant was controlled by a recessive gene, and it was different to the dense panicle genes which had been already cloned. Then by constructing the genetic segregated population and using the InDel markers, the small grain and dense panicle gene was mapped on the centromeric region of chromosome 5 between marker XP2628 and XP2631. In addition, the breeding application of the mutant was carried out on CMS lines and the stabile lines of high generations had finally been obtained.

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QTL Analysis on Grain Shape and Chalkiness of an Elite Maintainer Line Gang 46B in Hybrid Rice (Oryza sativa L.) GAO Fangyuan1,#, QIU Ling1,2,#, LU Xianjun1, REN Juansheng1, WU Xianting1, REN Guangjun1, ZENG Lihua2,* 2014, 28(3): 235-242.  DOI: 10.3969/j.issn.1001-7216.2014.03.003 Abstract ( )   PDF (694KB) ( )   To identify the genetic loci that control grain length, width, lengthwidth ratio, thickness, 100grain weight and the percentage of grains with chalkiness (GL, GW, LWR, GT, HGW, PGWC), we constructed an F2 population by crossing an elite  indica  maintainer line Gang 46B(G46B) with K1075, an introgression line of  G46B  derived from multibackcross with an US rice variety Lemont, with low percentage of chalkiness and narrow grain. All six investigated traits followed a continuous phenotypic distribution. Significantly positive or negative correlations were observed between all six traits, except the GL and GW. A genetic map was constructed with 30 simple sequence repeat (SSR) markers and a total of 14 QTLs for grain shape and chalkiness were detected by composite interval mapping (CIM). Among which, two were for GL (qGL2 and qGL5), two for GW (qGW5 and qGW8), five for LWR (qLWR2, qLWR5, qLWR6, qLWR8 and qLWR12), one for GT (qGT5), one for HGW (qHGW5), and three for PGWC (qPGWC5, qPGWC6 and qPGWC8). The G46B alleles contributed to increasing GW, GT, HGW and PGWC, while decreasing GL and LWR at all detected loci. The major QTLs controlling GW, GT, GW, PGWC and LWR were located in the same region RM17990RM18004RM18068 on chromosome 5, which was correspondent to the results identified in extreme phenotype plants using linkage and linkage disequilibrium.

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Association Analysis of Rice Grain Traits with SSR Markers TRAN Thi Thu Giang1,2, DANG Xiaojing1, LIU Qiangming1, ZHAO Kaiming1,3, WANG Hui1, HONG Delin1 ,* 2014, 28(3): 243-257.  DOI: 10.3969/j.issn.1001-7216.2014.03.004 Abstract ( )   PDF (2504KB) ( )   The genetic diversity, population structure and linkage disequilibrium of 540 rice varieties from different ecological regions were assessed with 262 pairs of simple sequence repeat (SSR) markers. The general linear model method in software of TASSEL was used to analyze the association between SSR markers and four grain traits. The main results were as follows: 1)the population was classified into seven subpopulations,with linkage disequilibrium at certain extent between SSR marker loci either intrachromosome or interchromosome within each subpopulation. 2)Among 45 grain lengthassociated SSR marker loci detected in all three years, RM5849 had the highest contribution rate (5286%), RM3766140 had the highest positive phenotypic effect, their carrier variety was Nongxiang 25. Among 7 grain widthassociated SSR marker loci, RM17 had the highest contribution rate (63.61%), RM17145 and RM573235 had the highest positive phenotypic effect, the  carrier varieties were Ningjinghui 338 and Zaoshirihuangdao, respectively. Among 11 grain thicknessassociated SSR marker loci, RM276 had the highest contribution rate (25.44%), RM508215 and RM7288205 had the highest positive phenotypic effect, carrier varieties were both Hongmangshajing. Among six 1000grain weightassociated SSR marker loci, RM17 had the highest contribution rate (32.17%), RM17145 had the highest positive phenotypic effect,  the carrier variety was Ningjinghui 338. Potential parental combinations for hybridization were predicted for improving grain traits by using the carrier varieties  screened.

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Effects of Different Paddy Field Drainage Degrees and Panicle Nitrogen Fertilizer Managements on Yield and Nitrogen Utilization of Different Nitrogen Efficiency Hybrid Rice ZHU Conghua1,2, SUN Yongjian1,2, YAN Fengjun1, JIANG Mingjin1, XU Hui1, ZHAO Jianhong1, MA Jun1,2,* 2014, 28(3): 258-266.  DOI: 10.3969/j.issn.1001-7216.2014.03.005 Abstract ( )   PDF (565KB) ( )   The effects of different paddy field drainage degrees including three soil water levels (0-20 cm soil volumetric water content was controlled at 53.60%±5.00%, 40.20%±5.00%, and 26.80%±5.00%, indicated by W1, W2, and W3 respectively) and three panicle nitrogen fertilizer managements (the panicle nitrogen fertilizer was applied on the 1st, 8th, and 15th day after rewatering, indicated by N1, N2, and N3 respectively) on dry matter productivity, nitrogen accumulation, translocation and utilization and yield of rice were investigated with nitrogen efficient variety(Dexiang 4103) and nitrogen inefficient variety(Yixiang 3724). The results showed that proper paddy field drainage degree could increase dry matter accumulation and translocation, nitrogen accumulation, translocation and utilization and yield of rice, and there are different effects for panicle nitrogen fertilizer management on yield and nitrogen utilization efficiency of different nitrogen efficiency rice. The dry matter accumulation and translocation, nitrogen accumulation, translocation and utilization of the nitrogen efficient variety were obviously higher than that of the nitrogen inefficient variety; the average yield of the nitrogen efficient variety was 1.12 t/hm2  higher than that of the nitrogen inefficient variety. Yield of nitrogen efficient variety and nitrogen inefficient variety had strongly positive correlation to dry matter accumulation and translocation and total nitrogen accumulation; yield of nitrogen efficient variety had strongly positive correlation to nitrogen translocation amount of stemsheath and yield of nitrogen inefficient variety had strongly positive correlation to nitrogen translocation amount of leaf and stemsheath. By applying the panicle nitrogen fertilizer on the 15th day after rewatering for nitrogen efficient variety (Dexiang 4103) under proper paddy field drainage condition treatment (W1) and applying the panicle nitrogen fertilizer on the 8th day after rewatering for nitrogen inefficient variety (Yixiang 3724) under proper paddy field drainage condition treatment (W2), nitrogen utilization efficiency and grain yield were all increased.

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Difference in Growth Duration and Utilization of Temperature and Solar Radiation Between indica and japonica Super Rice in the Lower Yangtze and Huaihe River Valley GONG Jinlong, XING Zhipeng, HU Yajie, ZHANG Hongcheng*, DAI Qigen, HUO Zhongyang, XU Ke, WEI Haiyan, GAO Hui 2014, 28(3): 267-276.  DOI: 10.3969/j.issn.1001-7216.2014.03.006 Abstract ( )   PDF (757KB) ( )   In order to reveal the difference in growth duration and utilization of temperature and solar radiation between indica and japonica super rice, a field experiment was conducted with five main representative super hybrid indica combinations  and five conventional japonica super rice varieties as materials in the ricewheat cropping areas. The main growth stages, dry matter accumulation, effective accumulated temperature, production efficiency of temperature, photosynthetically active radiation and solar energy utilization were analyzed systematically. Results showed that, the growth processes of japonica rice in the field were later than indica rice, 16.2 days later for rice maturity and harvest time. Grain filling stage of japonica rice was    significantly  longer than that of indica rice, with an increasing rate of 2582%. Compared with indica rice, japonica rice had strong adaptability to low temperature, which was beneficial for prolonging heading and grain filling appropriately, lengthening grain filling and growth duration, and increasing the utilization of temperature and solar radiation in late autumn. Japonica rice had significantly higher grain yield, biological yield, grain yield per day, and  growth duration, effective accumulated temperature, photosynthetically active radiation and solar energy utilization of the whole growth duration, and No. of days, effective accumulated temperature and photosynthetically active radiation of the major growth duration, and production efficiency of temperature from heading to maturity, and dry matter accumulation and solar energy utilization from jointing to maturity, while grain filling rate and production efficiency of temperature of the critical period from sowing to heading and the whole growth stages, and dry matter accumulation and solar energy utilization from sowing to jointing followed an opposite tendency accordingly. Correlation analysis showed that, grain yield of rice was found to be significantly positively associated with growth duration, grain yield per day and grain filling stage, while significantly negatively related to grain filling rate. Moreover, biological yield was highly significantly correlated with effective accumulated temperature, photosynthetically active radiation and solar energy utilization of the whole growth duration. However there was a insignificantly negative correlation between biological yield and production efficiency of temperature of the whole growth duration. Therefore, based on stabilizing and raising the utilization of temperature and solar radiation, grain yield per day and grain filling rate, the increment of effective accumulated temperature and photosynthetically active radiation through lengthening growth duration, especially for grain filling, could  increase periodic dry matter accumulation and biological yield, which was one of important approaches and characteristics of high productivity of japonica rice.

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Effects of Low Temperature on Grain Yield of Rice and Its Physiological Mechanism at the Booting Stage LI Jianling1,3, HUO Zhiguo2, WU Liji1, ZHU Qinghua1, HU Fei1，* 2014, 28(3): 277-288.  DOI: 10.3969/j.issn.1001-7216.2014.03.007 Abstract ( )   PDF (1634KB) ( )   Low temperature, especially occured at booting stage, would cause sterility of rice florets and became one of restricting factors of rice yield. In order to evaluate the effect of low temperature on yield of rice, and ascertain the low temperature warning indicator at  booting stage in South China, a controlled experiment was conducted with two cultivars, Tianyou 998 and Guinongzhan at four daily minimum temperature levels (18, 16, 14, and 12℃, diurnal temperature  range is 4℃,nutural air temperature as CK) with three durations (1, 3, and 5 d). The results showed that under the low temperature treatment during the booting stage of earlyseason rice, SOD and POD enzyme activities of the second leaf from top decreased, MDA content increased,SPAD value and photosynthetic rate declined, the percentage of pollen fertility and spikeletfertilized rate decreased. The number of grains per panicle, seedsetting rate, and 1000grain weight decreased, leading to a reduction in grain yield. When Tmin≤16℃ (3 d),the grain yield of Tianyou 998 and Guinongzhan significantly decreased, as well as development progress of grains and photosynthesis of the second leaf from top(P<0.05). The lower temperature and longer duration have a greater influence on the damage to rice. The coldtolerance of hybrid rice Tianyou 998 was weaker than the conventional rice Guinongzhan. The critical temperature for chilling damage was the daily lowest temperature 16℃ (3 days).

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Effects of Elevated CO2 Concentration, Nitrogen Fertilization, Planting Density and Foliar Zn Application on Rice Zn concentration and Bioavailability of Supper Rice Ⅱyou 084 under FACE Conditions ZHOU Sanni1, WANG Yunxia1, LAI Shangkun1, QI Yitao1, ZHU Jianguo2, YANG Lianxin1,*, WANG Yulong1,* 2014, 28(3): 289-296.  DOI: 10.3969/j.issn.1001-7216.2014.03.008 Abstract ( )   PDF (597KB) ( )   Elevated atmospheric CO2 concentration (\[CO2\]) increases rice yield, but little is known about effects of \[CO2\] on Zn bioavailability of edible parts of rice, as well as the agricultural practices which can improve micronutrient status of rice under high CO2 environment. By using a Free Air CO2 Enrichment (FACE) facility installed in paddy field, super rice Ⅱyou 084 were grown under two levels of CO2 concentration (ambient and 50% higher than ambient), N application (15 and 25 g/m2), planting density (16 and 24 hills/m2) and foliar Zn application (0% and 02% ZnSO4). At maturity, in milled and brown rice  Zn and phytate concentrations were analyzed, molar ratio of phytate to Zn were calculated. The results showed that compared to brown rice, milled rice had much lower Zn concentration, phytate concentration and molar ratio of phytate to Zn. Elevated \[CO2\] significantly decreased Zn concentration of milled and brown rice by 5% and 7%, respectively; Foliar Zn application significantly increased Zn concentration of milled and brown rice by 40% and 63%, respectively. However, the changes of N application and planting density had no effects on rice Zn concentration. Phytate concentrations of milled and brown rice were not affected by either CO2 or Zn treatments, but higher levels of N application and planting density significantly decreased phytate concentration of brown rice by 8% and 6%, respectively. The molar ratios of phytate to Zn in milled and brown rice were significantly increased at elevated \[CO2\]  in average of 6% and 7%, but decreased under foliar Zn application by 28% and 40%, respectively. The high levels of N application and planting density had little effects on molar ratio of phytate to Zn in milled rice, but in brown rice,it decreased by 10% and 7%. Our results indicate that the content and bioavailability of micronutrient Zn in edible part of rice Ⅱyou 084 will be lower under future high CO2 environment, but modification of agronomic practice such as ample N application, increase of planting density, especially foliar Zn application during grain filling stage can improve Zn nutrition of rice seeds to various degree.

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Influence of Flowering Characteristics on Spikelet Fertility under High Temperature ZHOU Jianxia, ZHANG Yuping, ZHU Defeng*, LIN Xianqing, XIANG Jing, CHEN Huizhe, HU Shengbo 2014, 28(3): 297-303.  DOI: 10.3969/j.issn.1001-7216.2014.03.009 Abstract ( )   PDF (1620KB) ( )   Ⅱyou 7954(medium heat sensitive) and Ⅱyou 7(heat tolerant) were subjected to heat stress (nine days or  three days,38℃) in phytotron at flowering stage to determine the influence of flowering characteristics on spikelet fertility under high temperature. The results showed that high temperature resulted in early flowering of Ⅱyou 7 and Ⅱyou 7954, and the former had obvious flowering peak, yet the latter had low or indistinct flowering peaks. There was obvious interaction between temperature and exposure duration for Ⅱyou 7954, and spikelet fertility reduced as thermal exposure prolonged. High temperature duration and temperature changing pattern, which had an effect on flowering habit in rice, affected spikelet fertility greatly.

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Haplotype Analysis of Rc Gene for Weedy Rice in Jiangsu Province LI Xiaoyan, QIANG Sheng, SONG Xiaoling, CAI Kun, DAI Weimin* 2014, 28(3): 304-313.  DOI: 10.3969/j.issn.1001-7216.2014.03.010 Abstract ( )   PDF (2468KB) ( )   Rc gene(about 6.4 kb) of 10 weedy rice samples from 10 cities in Jiangsu Province was sequenced and 166 Rc fulllength sequences of Oryza genus in literatures were reviewed. Colinear sequence analysis showed that Rc allele of Jiangsu weedy rice was wild genotype. Nucleotide polymorphism and haplotype analysis showed that Rc nucleotide polymorphism (π=0.19) and separation loci ratio (θw=0.28) of 10 weedy rice samples in Jiangsu Province were higher than that of 56 weedy rice samples from the United States (π=0.09; θw=0.07). Haplotype network and evolutionary phylogenetic analysis showed that Rc gene of weedy rice in Jiangsu was not directly derived from the revertant mutation of white cultivar. In addition, Rc gene sequences of Ludao from Lianyungang,Jiangsu and  Tangdao from Anhui were similar to that of cultivated rice, while genetically distant from weedy rice samples which has been leading to serious infestation in Jiangsu Province.

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Cloning and Characterization of Ty3/gypsy Retrotransposon in Chilo suppressalis (Lepidoptera: Pyralidae) LI Xiaohuan1, 2, ＃, LUO Guanghua2, ＃, ZHANG Zhichun2, LIU Baosheng2, FANG Jichao1, 2,* 2014, 28(3): 314-321.  DOI: 10.3969/j.issn.1001-7216.2014.03.011 Abstract ( )   PDF (2064KB) ( )   Transposable elements constitute a substantial fraction of host genomes. Ty3/gypsy retrotransposon, one group of LTR retrotransposons, is widespread in different species. By inversePCR and genome walking, a novel member of Ty3/gypsy retrotransposons was cloned from Chilo suppressalis. This new member of Ty3/gypsy retrotransposons was named CsuTy3 (GenBank accession No. KJ191261). The sequence is 4 934 bp in length and integrated into the “AACGT” target site duplications (TSDs) of the genome. There is a pair of noncompletely identical long terminal repeats (LTRs) in the CsuTy3 retrotransposon. The 5′LTR is 161 bp in length and the 3′LTR is 168 bp, sharing 935% similarities. A polypurine tract (PPT), 15 bp in length, is adjacent to the 3′LTR. There are three independent open reading frames (ORFs) in CsuTy3. The first ORF encodes a protein which is related to viral structural protein, termed GAG. The second encodes aspartic protease (AP). The third encodes a polyprotein including a reverse transcriptase (RT) which produces a cDNA copy of the transposon′s RNA, an RNase H (RH) which splits the DNARNA hybrid and an integrase (IN) which inserts the cDNA into the host′s genome. The southern hybridization indicated that there were many CsuTy3 copies in different C. suppressalis populations. Flanking PCR results showed that the CsuTy3 copy was inserted at the same locus in different populations. At this locus, all the individuals have the CsuTy3 copy insertion except some individuals from Deyang and Jiangjin populations. The CsuTy3 insertion ratio varied with field populations.

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Soilmicrobe Quantities and Enzyme Activities in Rhizosphere of Different Rice Varieties Fed by Brown Planthoppers SU Ting1, XU Hongxing2 , HAN Hailiang1, YANG Yajun2 , WANG Guiyue1,*, ZHENG Xusong2, LV Zhongxian2,* 2014, 28(3): 322-326.  DOI: 10.3969/j.issn.1001-7216.2014.03.012 Abstract ( )   PDF (360KB) ( )   The changes in pH value, soil enzyme activity and microbe density in the rhizosphere of potted rice plants fed by rice brown planthoppers (BPH), Nilaparvata lugens for different days were investigated on BPHsusceptible rice variety (TN1) and two resistant varieties (IR26 and IR36), for comparing the BPHfeedingassociated underground ecological systems among rice resistant varieties.The rhizosphere soil pH value of TN1 plants was significantly lower than that of BPHfree plants without significant deference among resistant varieties one day and five days after BPH feeding. The activities of sucrase and urease in rhizosphere did not differ among treatments one day after BPH feeding, while sucrase activity significantly increased and urease activity reduced in IR36 plant soil nine day after BPH feeding as compared with that in the BPHfree plant rhizosphere soil. The soil bacterial density in rhizosphere of IR36 significantly reduced only one day of BPH feeding, and fungal density in the rhizosphere of TN1 significantly reduced five days after BPH feeding, the soil actinomycete density in the rhizosphere of IR26 and IR36 significantly reduced. Multifactor analysis of variance showed that rice variety did not affect pH value and microbe population of rhizosphere soil and greatly affected the soil enzyme activity. Moreover, BPH feeding and feeding duration significantly affected pH value, soil enzyme activity and microbe density in the rhizosphere.

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Analysis on Yield and Quality of the Latematurity Medium japonica Rice Varieties Released in Jiangsu Province in the Last 30 Years ZHANG Honggen1, ZHU Guoyong1, FENG Zhiqiang1, XU Ming2, JI Jianan2, PEI Yan1, QIAN Kai1, TANG Shuzhu1,*, GU Minghong1 2014, 28(3): 327-334.  DOI: 10.3969/j.issn.1001-7216.2014.03.013 Abstract ( )   PDF (1671KB) ( )   In order to understand the breeding  situation and  trend of the latematurity medium  japonica  rice varieties in Jiangsu Province, the regional trial data and yield and quality of fortyfive latematurity medium japonica rice varieties released  in last thirty years were analyzed, which would provide a reference frame for the improvement and breeding of new varieties. The regional trial data showed that there were three development phases of yield of the latematurity medium japonica rice varieties and the representative varieties were Yanjing 2 with yield of over 500 kg/667m2 bred in 1983, Wuyujing 3 with yield of over 580 kg/667m2 bred in 1992, Wuyunjing 8 with yield of over 650 kg/667m2 bred in 1999, respectively. Yield of varieties bred after 2005 was fluctuated around 600 kg/667m2. Yield and quality of fortyfive latematurity medium  japonica rice varieties identified at Yangzhou in 2012 showed that  1) yield of varieties bred in different periods was all fluctuated from 500 kg/667m2 to 600 kg/667m2 and had not been developed significantly,  2) chalky grains of varieties had been improved significantly, but chalkiness and taste quality of varieties were still needed to be improved.