Evolutionary Genomics of Structural Variation in Asian Rice (Oryza sativa) Domestication

Structural variants (SVs) are a largely unstudied feature of plant genome evolution, despite the fact that SVs contribute substantially to phenotypes. In this study, we discovered SVs across a population sample of 347 high-coverage, resequenced genomes of Asian rice (Oryza sativa) and its wild ancestor (O. rufipogon). In addition to this short-read data set, we also inferred SVs from whole-genome assemblies and long-read data. Comparisons among data sets revealed different features of genome variability. For example, genome alignment identified a large (∼4.3 Mb) inversion in indica rice varieties relative to japonica varieties, and long-read analyses suggest that ∼9% of genes from the outgroup (O. longistaminata) are hemizygous. We focused, however, on the resequencing sample to investigate the population genomics of SVs. Clustering analyses with SVs recapitulated the rice cultivar groups that were also inferred from SNPs. However, the site-frequency spectrum of each SV type—which included inversions, duplications, deletions, translocations, and mobile element insertions—was skewed toward lower frequency variants than synonymous SNPs, suggesting that SVs may be predominantly deleterious. Among transposable elements, SINE and mariner insertions were found at especially low frequency. We also used SVs to study domestication by contrasting between rice and O. rufipogon. Cultivated genomes contained ∼25% more derived SVs and mobile element insertions than O. rufipogon, indicating that SVs contribute to the cost of domestication in rice. Peaks of SV divergence were enriched for known domestication genes, but we also detected hundreds of genes gained and lost during domestication, some of which were enriched for traits of agronomic interest.     

Photothermal Responses of Flowering in Rice (Oryza sativa)

Durations from sowing to panicle emergence in 16 diverse genotypesof rice (Oryza sativa L.) were recorded in 13 different photothermalregimes, comprising constant and diurnally alternating temperaturesbetween 16 and 32 °C and photoperiods between 10.5 and 15.0h d^–1—all provided by controlled-environment growthcabinets. In 11.5 h days and at sub-optimal temperatures, relationsbetween the rate of progress towards panicle emergence and meantemperature were linear in all genotypes, and amongst thesethe base temperature at that photoperiod varied between 6.6and 11.9 °C. In most cases progress was most rapid at 24–26°C, i.e. the optimum temperature was much cooler than expectedfrom previously published values of times to panicle emergencein a less extensive range of photothermal regimes. Only in threecultivars was it warmer than 28 °C, and in these there weresufficient data to establish that relations between rates ofprogress to panicle emergence and photoperiod in the diurnallyalternating temperature regime of 28–20 °C are alsolinear. Also, the responses of these three cultivars provideno evidence of any interaction between the effects of photoperiodand temperature. We conclude, then, that the model in whichrate of development is a linear function of both temperatureand photoperiod with no interaction, which has been shown tobe common to many other species, also applies to rice. Differencesamong genotypes in relative sensitivity of rate of progresstowards panicle emergence to both temperature and to photoperiodwere considerable; japonica cultivars tended to be more sensitiveto temperature and less sensitive to photoperiod than indicacultivars. Four indica cultivars bred and selected at The InternationalRice Research Institute (IRRI) in the Philippines did not differ(P > 0.10) in their relations between rate of progress towardspanicle emergence and sub-optimal temperatures in a daylengthof 11.5 h, but the optimum temperature for cv. IR 36 was appreciablywarmer than that for the cvs IR 5, IR 8 and IR 42. Oryza sativa, rice, flowering, temperature, photoperiod, photothermal responses     

水稻染色体G—带的研究

用改良的ASG法首次在籼稻(O.sativa subsp.indica)品种珍汕97和粳稻(O.subsp.iaponica)品种秀岭的有丝分裂染色体上显示了G-带,并作了相应的G-带核型分析。就同一材料来说,随着有丝分裂时期的推进,染色体上带纹数目逐渐减少。籼、粳亚种间相对应的同源染色体上G-带带纹特征彼此相似。讨论了水稻G-带带型与染色体不同区域分化的关系;G-带带型与籼、粳稻分歧的关系;以及G-显带的方法。     

Time-Course Association Mapping of the Grain-Filling Rate in Rice (Oryza sativa L.)

Detecting quantity trait locus (QTLs) and elite alleles that are associated with grain-filling rate (GFR) in rice is essential for promoting the utilization of hybrid japonica rice and improving rice yield. Ninety-five varieties including 58 landraces and 37 elite varieties from the core germplasm collection were genotyped with 263 simple sequence repeat (SSR) markers. The GFR of the 95 varieties was evaluated at five stages, 7, 14, 21, 28 and 35 days after flowering (DAF) both in 2011 and 2012. We found abundant phenotypic and genetic diversity in the studied population. A population structure analysis identified seven subpopulations. A linkage disequilibrium (LD) analysis indicated that the levels of LD ranged from 60.3 cM to 84.8 cM and artificial selection had enhanced the LD. A time-course association analysis detected 31 marker-GFR associations involving 24 SSR markers located on chromosomes 1, 2, 3, 4, 5, 6, 8, 9, 11 and 12 of rice at five stages. The elite alleles for high GFR at each stage were detected. Fifteen excellent parental combinations were predicted, and the best parental combination ‘Nannongjing62401×Laolaihong’ could theoretically increase 4.086 mg grain^-1 d^-1 at the five stages. Our results demonstrate that the time-course association mapping for GFR in rice could detect elite alleles at different filling stages and that these elite alleles could be used to improve the GFR via pyramiding breeding.     

缺磷条件下不同水稻品种磷素吸收特性的研究

缺磷条件下,供试粳稻品种植株的单株鲜重、干重、全磷含量和单株磷累积量等具有较大差异,从中筛选出磷高效品种TP309和优质8号,其中TP309较磷低效品种早88-1的全磷含量和单株磷累积量分别增加37.50%~40.00%和82.76%~102.00%。单株根数、平均根长和根系体积与单株磷素吸收量的相关程度较小,光合速率(Pn)、叶片可溶蛋白含量和叶片可溶性糖含量均表现为随着吸磷量的增加而不断增大。     

Rice (Oryza sativa) as a Remediation Tool for Nutrient Runoff

Hypereutrophication of U.S. surface waters is one of the leading causes of impairment for water quality. With nutrient criteria development and total maximum daily load (TMDL) issues looming for regulators, agricultural research is focusing on practices aimed at decreasing nutrient contributions to receiving aquatic ecosystems. This study examined the use of rice (Oryza sativa) for luxury uptake of nitrogen and phosphorus components associated with agricultural storm runoff. Mesocosms (379 L) planted with rice were exposed to two concentrations (5 and 10 mg/L) of nitrate, ammonia, and orthophosphorus. Results from these mesocosms were compared to unvegetated controls (also amended with 5 or 10 mg/L nitrate, ammonia, and orthophosphorus) to determine efficiency of rice in remediating nutrient runoff. Statistically significant differences in ammonia and nitrate retention of vegetated mesocosms amended with 5 mg/L versus vegetated mesocosms amended with 10 mg/L were noted after the first exposure. Although rice is a nutrient-dependent aquatic plant, this study suggests that more efficient mitigation is possible at lower inflow concentrations as opposed to higher inflow concentrations.     

水稻穗顶部退化基因PAA2的精细定位

鉴定和克隆水稻穗顶部退化突变体新基因,对研究小穗顶部退化的分子机制及克服育种和生产实践中因穗顶部退化引起的产量损失具有重要的理论和现实意义。本研究报道了一个来源于中花11的穗顶部退化突变体,暂命名为Panicle Apical Abortion 2(paa2)。该突变体的穗顶部小穗发育异常、退化,后期退化部分脱落,稻穗形成秃尖,使穗粒数减少。遗传分析表明该突变体受1个显性基因控制。利用群体分离分析法(BSA,bulked segregation analysis)将PAA2基因定位在2号染色体的长臂端L2-33和L2-50之间,物理距离为80 kb的范围内。该研究结果为PAA2基因的图位克隆奠定了基础。     

不同环境条件下稻米透明度的发育遗传分析

采用条件和非条件数量遗传分析方法,研究了不同环境条件下灿稻稻米透明度的发育遗传规律。稻米4个发育时期两年数据的分析结果表明,开花受精后第8～21天的灌浆中期和后期遗传效应表达对稻米透明度的影响最大。控制稻米透明度性状表现的三倍体胚乳核基因、细胞质基因和二倍体母体植株核基因效应在不同环境下存在着表达水平上的差异。条件遗传分析的结果还表明一些遗传效应存在着发育时期间断表达的现象。基因加性效应和细胞质效应以及相应的环境互作效应在稻米透明度性状发育中起着主要作用,选择可以取得良好的改良效果。浙南3号和1391等亲本可以提高后代的稻米透明度。     

水稻体细胞无性系变异

水稻体细胞无性系变异研究取得了很大进展 ,获得了大量抗病、抗逆、优质、矮杆等突变体。对这些突变体遗传分析表明 ,大多数突变性状由 1对或 2对基因控制。水稻体细胞无性系变异的发生与基因型、性状、继代时间、培养方式等有关 ,并具有内在的机制 ,点突变和反转录转座子插入可能是引起水稻无性系变异的两个重要原因。     

Coleoptile Senescence in Rice (Oryza sativa L.)

We investigated the cellular events associated with cell deathin the coleoptile of rice plants (Oryza sativa L.). Seeds germinatedunder submergence produced coleoptiles that were more elongatedthan those grown under aerobic conditions. Transfer of seedlingsto aerobic conditions was associated with coleoptile opening(i.e. splitting) due to death of specific cells in the sideof the organ. Another type of cell death occurred in the formationof lysigenous aerenchyma. Senescence of the coleoptile was alsonoted, during which discolouration of the chlorophyll and tissuebrowning were apparent. DNA fragmentation was observed by deoxynucleotidyltransferase-mediateddUTP nick end labelling (TUNEL) assay, and further confirmedby the appearance of oligonucleosomal DNA ladders in senescentcoleoptile cells. Two nucleases (Nuc-a and Nuc-b) were detectedby in-gel-assay from proteins isolated from coleoptiles. Nuc-a,commonly observed in three cell death phases required eitherCa²⁺or Mg²⁺, whereas Nuc-b which appeared during senescencerequired both Ca²⁺and Mg²⁺. Both nucleases were strongly inhibitedby Zn²⁺. Copyright 2000 Annals of Botany Company Aerenchyma, rice, cell death, coleoptile, fragmentation, nuclease, Oryza sativa, senescence, split, submergence, TUNEL     

粳稻米粒延伸性的QTL剖析

考察粳稻品种‘沈农265’和‘丽江新团黑谷’为亲本的F23,群体及双亲的米粒延伸率相关性状,并用该群体的分子连锁图谱进行QTL分析,共检测到8个与稻米延伸性有关的QTL,包括1个米粒型QTL、1个饭粒型QTL、2个延伸率QTL、3个膨胀率QTL和1个延伸指数QTL,分别位于第1、2、6和11染色体,单个QTL对性状表型贡献率在15．4％-37．5％之间。与其他研究结果比较表明,主效QTL在不同群体和不同环境中的重演性较好,这些QTL受水稻籼粳分化这一演化过程的影响较小。     

水稻产量及其构成因子对空气CO2浓度增高响应的QTL分析

自由空气CO2浓度增加设施（Free air carbon dioxide enrichment．FACE）使得实际地模拟未来植物生长所处的CO2浓度增加环境变为可能。FACE下．作物生长和产量发生不同程度的加速和提高,而分析作物产量因子对CO2浓度增加响应的遗传基础将有利于对CO2环境变化做出敏感响应的遗传特性的认识,有利于适合未来空气CO2浓度增加环境的高产品种的培育。以粳稻品种Asominori与籼稻品种IR24的杂交组合所衍生的染色体片段置换系（CSSLs）为材料进行田间试验,分别在FACE（约570umol CO2／mol）和正常大气（约370umol CO2／mol）下对籽粒产量及其构成因子等数量性状位点（QTL）进行了分析。结果表明,在FACE下,Asominori和IR24的有效穗数、穗粒数和单株籽粒产量均显著高于对照下的,并且FACE下,65个置换系的变幅范围均大于对照下的;在第1．2,4,6．7,9和12染色体上检测到LOD值在2．5—5．7范围内的控制上述产量性状的20个QTL．其中有3个可以同时在FACE和正常大气下检测到．其余的则只是在某一种CO2环境下检测到。此外,还检测到2个QTL（qFT12 and qGP4）存在着与环境的加性互作效应。可以推论．空气中CO2浓度的增加诱导了部分对CO2浓度敏感的QTL表达,控制水稻产量性状的QTL与CO2增加的环境发生了互作效应。预计利用分子标记辅助育种途径可以培育出适用于未来CO2浓度增加环境下的高产水稻品种。     

Linkage Disequilibrium in Salt Tolerant Genotypes of Rice (Oryza sativa L)

Genome wide linkage disequilibrium (LD) was investigated in a set of 32 genotypes representing salt tolerant improved varieties and landraces and six salt sensitive genotypes of rice with 64 microsatellite markers to identify the genomic regions that are associated with salt tolerance in rice. Out of 64 markers analyzed, 36% SSR pairs exhibited significant LD at 0.05. A few regions were identified as targets of selection in 10 chromosomes with high r ² values. The model-based groups from Bayesian clustering analysis are largely consistent with known pedigrees of the lines. The increased percentage of association of SSR loci in the improved varieties indicated the role of selection in linkage disequilibrium especially for salt tolerance. LD was extended as far as 100 cM in the present study. Most of the markers (43.8%) with significant LD values were observed in the genomic regions of reported QTL for salt tolerance in rice.     

授粉对水稻柱头蛋白质合成及钙离子富集的影响

应用非平衡等电聚焦和双向电泳技术,在不同水稻品种开花前后发育早期的雌蕊柱头中,发现两类5种电泳特性相似的主要多肽;在自交或杂交亲和授粉后并无明显变化;而在杂交不亲和授粉后,柱头中出现分子量为17．5kD,等电点为8．5的新带,可能与杂交不亲和授粉后的识别反应有关。在成熟柱头表面有大量钙离子富集,在幼嫩柱头表面却未发现;用成熟花粉强迫自交授粉,可诱导钙离子在幼嫩柱头表膜富集,说明钙离子的富集不但与柱头的成熟而且也与花粉的接触有关。