水稻RFLP连锁图谱的构建及控制小穗不育和花粉不育的QTL分析

用台中65(粳稻)/ARC10313(籼稻)的重组近交系(F10)构建了RFLP连锁图谱, 含113个分布均匀的标记. 作成的图谱覆盖全基因组, 全图总长1462.4 cM, 图中标记位置与所使用的参照图谱基本符合. 利用该重组自交家系材料与亲本台中65回交得到BF1家系, 用于对小穗不育和花粉不育的QTL分析, 检测出3个小穗不育和1个花粉不育QTL, 且有一     

水稻孕穗期耐热性QTLs分析

水稻籼粳亚种间杂种优势利用是提高水稻产量的重要途径。然而，异常高温或低温导致籼粳亚种间杂种育性下降是影响其优势利用的主要因素之一。本研究以USSR5（粳稻）/广解9号（籼稻）//USSR5回交群体为供试材料，构建了相应的分子连锁图谱，分别以高温处理下直接小穗育性及小穗育性热敏感指数为指标，对水稻孕穗期高温耐热性及其相对耐热性进行数量性状位点（QTLs）分析。结果表明，在第2、4和5染色体上检测到孕穗期耐热性相关的QTL各一个，对表型变异的解释率为6.4%~15.8%；在第4、8染色体上分别检测到与孕穗期相对耐热性相关的QTL，qhts-4和qhts-8，LOD值分别为3.81和2.86，对表型变异的解释率分别为16.8%和9.9%。对其进一步的上位性分析表明，有8条染色体的4对位点存在基因间互作，小穗育性耐热性除受主效QTL控制外，还受基因间互作及修饰基因的影响。     

Impact of Humidity on Temperature-Induced Grain Sterility in Rice (Oryza sativa L)

High temperature‐induced grain sterility in rice is becoming a serious problem in tropical rice‐growing ecosystems. We studied the mechanism of high temperature‐induced grain sterility of different rice (Oryza sativa L) cultivars at two relative humidity (RH) levels. Four varieties of Indica and Japonica rice were exposed to over 85 % RH and 60 % RH at 36/30 °C, 34/30 °C, 32/24 °C and 30/24 °C day/night air temperatures from late booting to maturity inside sunlit phytotrons. Increasing both air temperature and RH significantly increased spikelet sterility while high temperature‐induced sterility decreased significantly with decreasing RH. Neither Indica nor Japonica rice types were superior to the other in the response of their spikelets to increased air temperature and RH. Increased spikelet sterility was due to increased pollen grain sterility which reduced deposition of viable pollen grains on stigma. Reduction in sterility with decreased RH was more due to decreased spikelet temperature than to air temperature. Thus the impact of RH should be considered when interpreting the effect of high temperature on grain sterility. Spikelet fertility was curvilinearly related to spikelet temperature. Grain sterility increased when spikelet temperature increased over 30 °C while it became completely sterile at 36 °C. The ability of a variety to decrease its spikelet temperature with decreasing RH could be considered as avoidance while the variability in spikelet sterility among varieties at a given spikelet temperature could be considered as true tolerance.     

一个新的水稻花粉半不育性位点的定位分析

利用一套以籼稻珍汕97B为背景的粳稻日本晴染色体片段代换系,鉴定发现1个半不育的代换系。全基因组基因型分析表明,该代换系仅含3个粳稻导入片段,而其他遗传背景与珍汕97B相同。在湖北武汉和海南分别种植其衍生的F₂和F₃分离群体,采用单标记分析和区间作图法分析花粉育性和小穗育性的数量性状位点(QTL),结果表明,该代换系的半不育性是第2染色体上的粳稻导入片段引起的,该片段RM262~RM475区间存在1个新的影响花粉育性的QTL,其贡献率为13.9%。研究结果将为进一步精细定位水稻育性QTL以及鉴定相关功能基因提供重要的试验基础。     

东乡野生稻细胞质雄性不育育性恢复的遗传研究

应用协青早A／（协青早B／／东乡野生稻／协青早B）BC1F5、中9A／（协青早B／／东乡野生稻／协青早B）BC1F5、协青早A/（协青早B／／协青早B／东乡野生稻）Be1F5和中9A／（协青早B／／协青早B／东乡野生稻）BC1F5四套测交群体,分析测交群体的育性表现及研究东乡野稻细胞质雄性不育育性恢复的遗传。结果表明：东乡野生稻对矮败（CMS—DA）和印尼水田谷败育（CMS—IA）的育性恢复表现为质量一数量性状,结合群体遗传结构、育性分布和东乡野生稻的偏粳性及广亲和性,可知东乡野生稻对CMS—DA和CMS—IA的恢复性由1对或2对恢复基因控制。     

Identification of four genes for stable hybrid sterility and an epistatic QTL from a cross between Oryza sativa and Oryza glaberrima

To further understand the nature of hybrid sterility between Oryza sativa and Oryza glaberrima, quantitative trait loci (QTL) controlling hybrid sterility between the two cultivated rice species were detected in BC₁F₁ and advanced backcross populations. A genetic map was constructed using the BC₁F₁ population derived from a cross between WAB450-16, an O. sativa cultivar, and CG14, an O. glaberrima cultivar. Seven main-effect QTLs for pollen and spikelet sterility were detected in the BC₁F₁. Forty-four sterility NILs (BC₆F₁) were developed via successive backcrosses using pollen sterility plants as female and WAB450-16 as the recurrent parent. Seven NILs, in which the target QTL regions were heterozygous while the other QTL regions as well as most of the reminder of the genome were homozygous for the WAB450-16 allele, were selected as the QTL identification materials. BC₇F₁ for the seven NILs showed a continuous variation in pollen and spikelet fertility. The four identified pollen sterility QTLs were located one each on chromosomes 1, 3, 7 and 7. Pollen sterility loci qSS-3 and qSS-7a were on chromosomes 3 and 7, respectively, which coincides with the previously identified S19, and S20, while loci qSS-1 and qSS-7b on chromosomes 1 and 7L appear distinct from all previously reported loci. An epistatic interaction controlling the hybrid sterility was detected between qSS-1 and qSS-7a.     

水稻光温敏雄性核不育系45S不育基因的分子定位

水稻光温敏雄性核不育系45S的育性转换受光长和温度的互补作用调控。对45S（Oryza sativa L．ssp．indica）WC169组合的F2,BC1F1和F2和BC2F2进行不育基因的遗传分析,结果表明45S的育性是由1对隐性不育基因控制。利用SSR分子标记技术,结合BSA和RCA法,以45S与YC169的杂交F2和BC2F2作为分离群体,对不育基因进行定位,结果发现不育基因（暂命名为GTMS）位于RM6378和RM12847之间,遗传距离分别为8．5cM和8．2cM。     

Quantitative trait loci for stay-greenness and agronomic traits provide new insights into chlorophyll homeostasis and nitrogen use in rice

Functional stay-green (FSG) is characterized by delayed senescence during the reproductive stage in rice. A recombinant inbred line (RIL) population derived from ‘Pusa 677’/‘PSG16’ was used to study the genetics of FSG in a rice mutant, ‘PSG16’. The RILs exhibited significant variation under two N regimes for agro-morphological traits, leaf chlorophyll content, flowering time, yield components and nitrogen (N) use. Using a genome wide linkage map spanning 1910.8 cM with 104 polymorphic markers, we have mapped six major quantitative trait loci (QTLs). One robust QTL on chromosome 1 was associated consistently across different N regimes with chlorophyll content and flowering time. The QTL on chromosome 7 was associated with grain number, whereas the QTL on chromosome 6 was found related to N harvest index and spikelet fertility. Although ‘PSG16’ showed a clear advantage in grain yield as well as having better N assimilation, we could not establish a direct genetic relationship SG trait and N use. Based on the QTL information, FSG trait of ‘PSG16’ could be useful in deciphering multiple stress responses in rice.     

Effects of Increasing Temperatures on Spikelet Fertility in Different Rice Cultivars based on Temperature Gradient Chamber Experiments

Spikelet sterility in rice (Oryza sativa L.) induced by high temperatures is a major concern given global warming predictions. We studied differences among eight rice cultivars in spikelet fertility at five different temperature levels in temperature gradient chamber (TGC) experiments. Six japonica and two indica cultivars were exposed to high‐temperature gradients in TGCs during the 2005 flowering season. Spikelet sterility increased with temperature in TGCs and differed among cultivars because of both variations in temperature tolerance and timing of heading. The correlation between spikelet fertility of individual panicles and both air temperature and panicle temperature during flowering was analyzed to compare tolerances among cultivars. The temperature (T₇₅) at which spikelet fertility was 75 % of maximum ranged from 34 to 39 °C air temperature and differed significantly among cultivars. Indica varieties had higher T₇₅ values than japonica varieties. The T₇₅ values based on panicle temperature also differed among cultivars, but the difference between indica and japonica varieties were less significant. We concluded that the higher temperature tolerances of indica cultivars in our experiments could be attributed to lower spikelet temperatures, and cultivars with similar spikelet temperatures still had different heat tolerances due to differences in pollination ability.     

Responses of spikelet fertility to air,spikelet, and panicle temperatures and vapor pressure deficit in rice

High temperature-induced spikelet sterility is expected to become a major factor reducing rice yield under the future climate conditions. To examine the responses of spikelet sterility to air temperature, humidity, and temperatures of panicle and spikelet, two japonica rice cultivars in different maturity groups were exposed to four sets of different temperature conditions from initial heading stage in 2013 and 2014. The temperature conditions included ambient temperature (AT), AT + 1.5°C, AT + 3.0°C, and AT + 5.0°C. Spikelet fertility showed a wide range of variation from 100 to 4.6% depending on temperature treatments. The ridge regression revealed that not only air temperature but also vapor pressure deficit (VPD) was negatively associated with spikelet fertility. The spikelet fertility was well fitted to logistic equations not only of air temperature, spikelet internal temperature, and panicle surface temperature but also of VPD. No clear difference in the accuracy of sterility estimation was observed for models that use air temperature, panicle or spikelet temperature as inputs. In contrast, the logistic equation model that uses both air temperature and VPD as independent variables had better accuracy in predicting spikelet sterility. These results suggested that spikelet or panicle temperature would be no better predictor for high temperature-induced spikelet sterility than air temperature. Therefore, further study is merited to verify the VPD effects on spikelet sterility under high temperature conditions.     

利用籼稻资源中的“隐蔽有利基因”提高粳稻苗期耐冷性

苗期低温是制约东北水稻生产的一个重要逆境因子,培育苗期耐低温的水稻品种是解决这一问题最有效的途径之一。以粳型恢复系C418为轮回亲本与7个来自不同地域的供体杂交和回交培育BC₂F₂群体，利用沈阳春季田间自然低温进行耐冷筛选，并对入选的177个苗期耐冷导入系后代在自然和人工气候室两种不同强度的低温胁迫条件下的苗期表现进行评价。结果表明，虽然供体亲本均为苗期耐冷性弱的品种，但在全部回交后代中均出现耐冷的超亲分离，表明这些供体亲本均携带对耐冷性有利的“隐蔽基因”, 不过组合间的耐冷性选择效率存在较大差异。以人工气候室低温胁迫后的存活率为耐冷性鉴定指标，导入系之间出现广泛分离，田间筛选入选比例高的BG300和中413导入系群体中出现存活率高的导入系后代比例较其它群体高，说明在同一轮回亲本遗传背景下回交后代的耐冷水平取决于供体。在室内严格低温条件下表现为强耐冷性的导入系在相对温和的自然低温胁迫下表现为苗高、苗干重较非胁迫条件生长抑制不明显，且能够促进根系生长。结果表明，大量的耐冷有利基因以“隐蔽”的形式存于水稻种质资源中。通过利用来源广泛的种质资源作为供体进行回交育种，对回交后代进行严格耐冷筛选，可有效选育高产耐冷水稻品种和有价值的育种材料。     

应用SSR标记定位水稻再生力和再生产量及其构成的QTL

用两个籼稻品种明恢86和佳辐占为亲本建立的F2群体及相应的SSR分子标记连锁图,对水稻再生力和再生产量及其构成的基因进行了定位。结果定位了影响水稻再生力(再生穗数)的1个QTL,影响再生产量的1个QTL和影响产量构成的2个QTL,各QTL的加性效应均来自亲本明恢86,起增效作用。影响水稻再生穗数、结实率和单株产量的QTL位于同一染色体的相同区段上,且加性效应方向相同,这很好地解释了再生穗数与单株产量、结实率呈极显著正相关的关系。     

Increasing biodiversity of irrigated rice in Africa by interspecific crossing of Oryza glaberrima (Steud.) × O. sativa indica (L.)

The African rice Oryza glaberrima, traditionally cultivated since more than 3.500 years, is of poor agronomic performance but resistant/tolerant to various stresses and diseases. The introduction of these characters into O. sativa cultivars is difficult since crossing barriers cause spikelet sterility in F1. Backcrossing can restore fertility and recently facilitated the development of fertile O. glaberrima × O. sativa ssp. japonica hybrid progenies for rain fed systems. With the objective to gain access to African rice germplasm for improvement of irrigated rice, crosses were performed with eighteen O. glaberrima and twenty O. sativa ssp. indica accessions. In total about one hundred F1-hybrid grains were obtained. The F1 plants were all completely sterile and backcrossing (BC) to O. sativa was performed in order to restore spikelet fertility. Monitoring of Tog5681 × IR64 hybrid progenies under field conditions revealed a broad genetic diversity within the BC₁ and BC₂ populations. Some BC₁ and BC₂ progeny plants outperformed the O. sativa parent, indicating that the heterozygocity level and complementary gene action after two backcrosses are still sufficient to positively influence plant vigor. Spikelet fertility of progenies was highly variable, but almost complete fertility was already observed within the BC₁F₂ population. High spikelet fertility was preserved in one out of two analyzed BC₁F₃ families and inmost of the BC₂F₃ families. The ability to restore spikelet fertility within few generations and the potential of the genetic diversity present in interspecific progenies facilitates the development of plant types specifically designed for the African irrigated and lowland environment. This revised version was published online in August 2006 with corrections to the Cover Date.     

西藏三联小穗小麦中三联小穗性状控制基因的SSR分子标记定位

西藏三联小穗小麦是中国西藏地区一种独特的小麦地方品种,拥有特殊的三联小穗性状,超多的小穗数和小花数。分子定位控制三联小穗基因的基因座,发掘与之紧密连锁的分子标记,可为小麦高产育种提供分子标记辅助选择工具。本研究利用西藏三联小穗小麦的衍生系TTSW-5与普通穗型小麦,间3和川麦55,分别构建F2群体,成熟后进行穗部性状的表型分析和SSR基因型鉴定。性状表型遗传分析表明,西藏三联小穗小麦的三联小穗性状由两个独立遗传的隐性基因控制;通过SSR标记鉴定来自TTSW-5/间3组合的F2群体,在2A染色体上检测到1个与三联小穗性状相关的QTL,定位于SSR标记Xgwm275和Xgwm122之间,两标记间的遗传距离为6.6cM,该QTL的LOD值为6.19,可解释的表型变异值为33.1%,初步命名为qTS2A-1。我们推测qTS2A-1可能是控制三联小穗性状相关的主效QTL,SSR标记Xgwm275和Xgwm122可能可用于三联小穗性状的辅助选择。     

东乡野生稻抗褐飞虱QTL分析

野生稻是水稻抗褐飞虱基因的重要种质资源。应用东乡野生稻与栽培稻协青早B构建的2套材料,开展水稻抗褐飞虱基因鉴定研究。先以协青早B//协青早B/东乡野生稻BC₁F₅群体为材料,应用褐飞虱田间种群进行抗虫鉴定,检测到2个抗褐飞虱QTL,其中,qBph2位于水稻第2染色体RM29–RG157区间,东乡野生稻等位基因可降低死苗率22.2%;qBph7位于第7染色体RM11–RM234区间,东乡野生稻等位基因可降低死苗率43.7%。进一步以协青早B为轮回亲本,构建了BC₃F₃群体,应用褐飞虱生物型I、II和III进行抗虫鉴定,QTL分析表明qBph2抗褐飞虱生物型I和II,qBph7抗褐飞虱生物型I和III。这2个QTL对培育抗褐飞虱水稻品种具有重要应用价值。     

不同环境条件下稻米组氨酸和精氨酸的胚乳和母体植株QTL分析

利用汕优63重组自交系与双亲回交产生的BC1F1和BC2F1群体,采用新发展的包括环境互作效应在内的多遗传体系QTL作图方法和基因定位软件,对稻米两种半必需氨基酸（组氨酸和精氨酸）进行三倍体胚乳和二倍体母体植株等不同遗传体系的QTL定位分析。共检测到10个控制组氨酸含量的QTL以及8个控制精氨酸含量的QTL。全部QTL均具有极显著的三倍体胚乳和二倍体母体植株基因的加性主效应,其中4个QTL具有显著或极显著的三倍体胚乳显性主效应,7个QTL还具有明显的环境互作效应。     

Fine mapping of <Emphasis Type="Italic">Rf3</Emphasis> and <Emphasis Type="Italic">Rf4</Emphasis> fertility restorer loci of WA-CMS of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) and validation of the developed marker system for identification of restorer lines

The Wild Abortive (WA) system is the major cytoplasmic male sterility (CMS) source for hybrid rice production in indica rice and its fertility restoration is reported to be controlled by two major loci viz. Rf3 on chromosome 1 and Rf4 on chromosome 10. With the availability of the rice genome sequence, an attempt was made to fine map, develop candidate gene based markers for Rf3 and Rf4 and validate the developed marker system in a set of known restorer lines. Using polymorphic markers developed from microsatellite markers and candidate gene based markers from Rf3 and Rf4 loci, local linkage maps were constructed in two mapping populations of ~1,500 F₂ progeny from KRH2 (IR58025A/KMR3R) and DRRH2 (IR68897A/DR714-1-2R) hybrids. QTLs and their interactions for fertility restoration in Rf3 and Rf4 loci were identified. The identified QTL in both mapping populations together explained 66–72 % of the phenotypic variance of the trait suggesting their utility in developing a marker system for identification of fertility restorers for WA-CMS. Sequence comparison of the two candidate genes from the Rf3 and Rf4 regions in male sterile (A) and restorer (R) lines showed 2–3 bp indels and a few substitutions in the Rf3 region and indels of 327 and 106 bp in the Rf4 region respectively. The marker system identified in the present study was validated in 212 restorers and 34 maintainers along with earlier reported markers for fertility restoration of WA-CMS. Together DRCG-RF4-14 and DRCG-RF4-8 for the Rf4 locus and DRRM-RF3-5/DRRM-RF3-10 for the Rf3 locus showed a maximum efficiency of 92 % for identification of restorers.     

棉属野生种克劳茨基棉第7染色体上马克隆值QTL的挖掘与定位

为了深入挖掘和利用棉属野生种克劳茨基棉(Gossypium klotzschianum)的优异等位基因，构建了一个(陆地棉泗棉2号×克劳茨基棉)×泗棉2号的BC₁F₂群体，对纤维品质性状初步定位，单标记相关分析表明位于第7染色体上的SSR标记NAU1362与马克隆值表现极显著相关。进一步选择在第7染色体上含有克劳茨基棉渐渗片段的BC₁F₂单株与轮回亲本泗棉2号回交，构建BC₂F₃和BC₂F₄分离群体，通过两年的田间重复试验验证该QTL的位置与效应。结果表明，该QTL (qFMIC-7-1)在BC₂F₃、BC₂F₄世代均被检测到，位于相同的标记区间，分别可以解释9.0%、8.8%的表型变异，增效基因来源于野生种克劳茨基棉，与BC₁F₂群体定位结果基本一致。同时在第7染色体上检测到另一马克隆值QTL (qFMIC-7-2)，同样在BC₂F₃、BC₂F₄两个世代均能够被检测到，分别可以解释3.7%、4.7%的表型变异，但增效基因均来源于泗棉2号。     

Dissection of genetic architecture for oil content in soybean seed using two backcross populations

Soybean seed oil was valued in foods, animal feed and some industrial applications. Molecular marker‐assisted selection (MAS) for high‐oil‐content cultivars was an important method for soybean breeders. The objective of this study was to identify quantitative trait loci (QTL) and epistatic QTL underlying the seed oil content of soybeans across two backcross (BC) populations (with one common male parent ‘Dongnong47’) and two different environments. Two molecular genetic maps were constructed. They encompassed 1046.8 cM [with an average distance of 6.75 cM in the ‘Dongnong47’  ×  ‘Jiyu89’ (DJ) population] and 846.10 cM [with an average distance of 5.76 cM in the ‘Dongnong47’  ×  ‘Zaoshu18’ (DZ) population]. Nine and seven QTL were identified to be associated with oil content in the DJ and DZ populations, respectively. The phenotypic variation explained by most of the QTL was usually less than 10%. Among the identified QTL, those stable ones across multiple environments and populations often had stronger additive effects. In addition, three stable QTL in the DZ populations were identified in the similar genomic region of the three QTL in the DJ population [qDJE and qDZE‐1 were located near Satt151 of Chromosome 15 (Chr15), qDJA1 and qDZA1 were located near Satt200 of Chr15 (LG A1), and qDJD2‐1 and qDZD2‐1 were located near Sat365 of Chr17]. In conclusion, MAS will be able more effectively to combine beneficial alleles of the different donors to design new genotypes with higher soybean seed oil content using the BC populations.