水稻剑叶形态QTL定位及候选基因分析

叶片是水稻的重要器官,是进行光合作用的主要场所,而叶长、叶宽、叶面积等剑叶形态相关性状能够直接影响叶片的光合效率.研究水稻剑叶形态相关性状的QTL(quantitative trait loci)位点,并通过遗传育种的手段对水稻剑叶形态进行改良,可以为培育理想株型的水稻提供理论依据.本研究以籼稻品种华占(HZ)为父本,...     

水稻抗稻瘟病近等基因系的cDNA微阵列分析

应用cDNA微阵列对来源于中156／谷梅2号重组自交系的水稻抗稻瘟病近等基因系G205和G71的稻瘟病菌胁迫基因表达谱进行了分析,发现有3个cDNA克隆的表达仅在抗病基因系G205接种病原菌12h后受到诱导,其中两个为功能已知基因,另一个为功能未知的新基因。另有35个差异表达克隆在两个近等基因系中均检测到,其中17个克隆的表达在G205和G71均受到病原菌的诱导,另外18个克隆的表达则在G205和G71均受到病原菌原抑制。序列分析表明,这些稻瘟病菌应答基因分别与防卫反应,信号传递,逆境胁迫和光合作用及糖代谢等功能相关,为植物抗病机制提供了相关信息。另外,Northern还证实了编码富含甘氨酸蛋白基因（Glycinerich protein Grp)的表达受稻瘟病病原菌的诱导,是一个稻瘟病诱导相关基因。     

水稻品种USSR5早熟性的遗传分析

USSR5为极早熟的前苏联品种,以抽穗期近等基因系和抽穗期QTL近等基因系为测验品种,对USSR5的抽穗期基因型进行分析,表明USSR5携带了非感光基因e1、无感光功能的Se-1e基因、感光抑制基因i-Se-1和显性早熟基因Ef-1,从而使它表现极早熟的特性。此外,本研究调查了USSR5和N22的BC1F1和F2群体的抽穗期,利用WindowsQTLCartographer1.13a软件,采用复合区间作图法,在全基因组范围内,分析了南京夏季正常日照条件下2个群体的抽穗期QTL,在USSR5/N22//USSR5BC1F1群体,共检测到2个位点,分别位于第7、8染色体上,其LOD值分别是6.11和2.91,对表型总变异的解释率分别为27.38%和11.15%,2个位点上来自USSR5的等位基因均提早抽穗。在USSR5/N22F2群体,共检测到5个位点,分别位于第1、2、7、9、10染色体上。5个位点LOD值介于3.02～8.4,对表型总变异的解释率分别为4.07%和15.41%。除qHd-9外,其余控制抽穗期的4个基因位点上提早抽穗的等位基因均来源于USSR5。比较分析发现效应较大的qHd-7即是Hd4(E1),USSR5在该位点上携带非感光基因hd4(e1)。尽管本研究定位的其它抽穗期QTL和已知抽穗期基因之间尚不能一一对应,但在早熟性水稻品种选育中,USSR5将可作为良好的基因源加以利用。     

水稻雄性不育恢复系明恢63的感光基因分析

水稻恢复系明恢63是中国应用面积最大、利用最广泛的恢复系。利用抽穗期感光性近等基因系EG0～EG7及ER～LR对明恢63进行的分析表明,明恢63在E1、E2、E3位点分别带有E1、e3、E3基因,在Se-1位点带无感光功能的Se-1^e基因。进一步用抽穗期QTL近等基因系NIL(Hd1)和NIL（Hd4)进行的研究表明,明恢63带有显性感光基因E1和无感光功能的Se-1^e基因,并推测明恢63带有能抑制E,基因表达的隐性抑制基因。认为籼型杂交稻抽穗期受不育系和恢复系感光基因及感光性抑制基因的共同作用。初步讨论了明恢63广适性的遗传基础。     

甜菜夜蛾抗高效氯氟氰菊酯近等基因系的RAPD标记

用随机引物扩增的方法对近等基因系抗高效氯氟氰菊酯种群和敏感种群甜菜夜蛾进行比较分析,从160条引物中筛选,引物S42(GGACCCAACC)和OPH13(GACGCCACAC)在抗性种群中扩增分别得到了1条特异扩增带,带型清晰、重复性较好,所以S42和OPH13产生的两条特异扩增带是分别与甜菜夜蛾对高效氯氟氰菊酯抗性相关的RAPD分子标记。     

水稻白叶枯病新抗源Y238的鉴定及其近等基因系培育

从269份普通野生稻中鉴定出一个高抗白叶枯病的新抗源,编号为Y238.通过多茵系鉴定、抗谱分析及与目前国际上已知基因比较,证明该新抗源含有一个新基因,暂命名为WBB2.对JG30/Y238杂交后代成株期接种鉴定、遗传分析表明,WBB2为完全显性基因.通过杂交和回交,已将WBB2导入栽培稻中构建近等基因系.     

基因聚合提高了水稻对白叶枯病的抗性

研究了含有单个抗性基因的水稻近等基因系和抗性基因聚合品系对浙江省白叶枯病菌４个主要小种的抗性,单个基因对这些小种的抗性均不高,对新近流行的小种大多感病;基因聚合品系对这些小种的抗性普遍提高,说明基因聚合是培育具有持久抗性品种的有效策略。     

水稻株高和抽穗期基因的定位和分离

利用241个重组自交系构成的群体,对水稻（Oryza sativaL.)株高和抽穗期进行基因定位,三年共定位到4个抽穗期的数量性状基因（QTLs)和4个株高OTLs,其中位于第7染色体C1023-R1440区间的QTL3年均可检测到,且效应大,同时影响株高和抽穗期。为了区分这个区间的QTL是一因多效还是紧密连锁的两个QTLs,从自交系群体里选取QTL区间来自明恢63,其他遗传背景与珍汕97高度相似的自交系RⅡ50,与珍汕97回交,获得含有363个单株的近等基因系BC1F2群体。考察株高和抽穗期。两个性状在群体里表现为双峰分布,它们的分离比符合期望的单基因盂德尔遗传分离比,BC1E2群体单株的株高和抽穗期基本表现为矮秆早抽穗,高秆迟抽穗,但是,6个单株表现相反的情况,以上结果证明,QTL能够作为盂德尔因子进行研究,在BC1F2群体里,株高和抽穗期是由单个基因控制的,第7染色体上是两个紧密连锁的基因分别控制株高和抽穗期。     

小麦Mlo及NBS-LRR类抗病基因同源序列的分离与鉴定

根据GenBank中公布的大麦白粉病抗性控制基因Mlo cDNA序列及一个来源于栽培一粒小麦(Triticum monococcum L.)的假定抗病基因序列分别设计引物,以携带小麦抗白粉病基因的近等基因系为材料进行RT-PCR筛选.结果获得两个表达基因的cDNA克隆.其中一个与大麦白粉病抗性控制基因Mlo的同源性达83%.另一个为非通读序列,含有两个可能的开放阅读框,分别包含抗病基因NBS保守结构域2和3以及与水稻抗稻瘟病基因Pib蛋白末端相似的13个LRR区域,推测该序列属于NBS-LRR类.白粉菌诱导前后,该片段RT-PCR扩增产物存在差异,表明该片段可能与小麦抗病性相关.利用\"中国春\"缺体-四体系,将该NBS-LRR类序列定位在小麦1D染色体上.     

水稻广亲和基因（WCG）近等基因系的随机引物PCR分析

应用随机引物ＰＣＲ（ＲａｎｄｏｍＰｒｉｍｅｒＰＣＲ）技术分别在水稻广亲和基因（ＷＣＧ）的近等基因系和色素原基因（Ｃ）的分离群体库中寻找与ＷＣＧ和Ｃ基因连锁的分子标记。对于ＷＣＧ近等基因系,在２２６个随机引物中初筛到２２个显示多态性片段的引物。根据理论值计算,在２２个多态性片段中预期有２０个与ＷＣＧ连锁。在这些连锁标记中距ＷＣＧ最近的可达０．５ｃＭ。同样在分离群体库的筛选中有１０个扩增产物与Ｃ基因连锁。     

Evaluation of a leaf area prediction model proposed for sunflower

Six leaf samplings were conducted in two sunflower (Helianthus annuus L.) hybrids during the 2006 growing season in order to evaluate a simple model proposed for leaf area (LA) estimation. A total of 144 leaves were processed using an image analysis system and LA, maximum leaf width (W) [cm], and midvein length (L) [cm] were measured. Also, LA was estimated using the model proposed by Rouphael et al. (2007). Measured LA was exponentially related with L and W, and the W-LA relationships showed higher r ². Estimated LA was strongly and exponentially related with L. Strong, linear relationships with high r ² between estimated and measured LA confirmed the high predictability of the proposed model.     

Leaf area estimation in a sugar beet cultivar by linear models

An indirect method of leaf area measurement for Rizor sugar beet cultivar was tested. Leaves were sampled during two growing seasons in a Randomised Complete Block Design experiment. For 2002 samplings, leaf area [cm²] was linearly correlated with maximum leaf width [cm] using all leaf samples (r ² = 0.83, p < 0.001) or using the means of the 8 sampling occasions (r ² = 0.97, p < 0.001). Correlations between leaf area and leaf mid vein length [cm] were weaker (r ² = 0.75, p < 0.001 and r ² = 0.93, p < 0. 001, respectively). For 2003 samplings, the area estimated by the equations was highly correlated to the measured leaf area.     

Nitrogen fertilization effects on leaf morphology and evaluation of leaf area and leaf area index prediction models in sugar beet

In a two-year experiment (2002–2003), five N application rates [0, 60, 120, 180, and 240 kg(N) ha⁻¹, marked N₀, N₆₀, N₁₂₀, N₁₈₀, and N₂₄₀, respectively] were applied to sugar beet cv. Rizor arranged in a Randomized Complete Block design with six replications. Leaf shape parameters [leaf area (LA), maximum length (L), maximum width (W), average radial (AR), elongation (EL), and shape factor (SF)] were determined using an image analysis system, and leaf area index (LAI) was non-destructively measured every two weeks, from early August till mid-September (four times). Years, samplings, and their interaction had significant effects on the determined parameters. Fertilization at the highest dose (N₂₄₀) increased L and sampling×fertilization interaction had significant effects on LA, L, W, and SF. For this interaction, W was the best-correlated parameter with LA and LAI meaning that W is a good predictor of these parameters. Two proposed models for LA estimation were tested. The model based on both leaf dimensions [LA = 0.5083 (L×W) + 31.928] predicted LA better than that using only W (LA = 21.686 W − 112.88). Instrumentally measured LAI was highly correlated with predicted LAI values derived from a quadratic function [LAI = −0.00001 (LA)² + 0.0327 LA − 2.0413]. Thus, both LA and LAI can be reliably predicted non-destructively by using easily applied functions based on leaf dimensions (L, W) and LA estimations, respectively.     

Mapping of QTLs conferring resistance to bacterial leaf streak in rice

A large F₂ and a RI population were separately derived from a cross between two indica rice varieties, one of which was highly resistant to bacterial leaf streak (BLS) and the other highly susceptible. Following artificial inoculation of the RI population and over 2 years of testing, 11 QTLs were mapped by composite interval mapping (CIM) on six chromosomes. Six of the QTLs were detected in both seasons. Eight of the QTLs were significant following stepwise regression analysis, and of these, 5 with the largest effects were significant in both seasons. The detected QTLs explained 84.6% of the genetic variation in 1997. Bulked segregant analysis (BSA) of the extremes of the F₂ population identified 3 QTLs of large effect. The 3 QTLs were dentical to 3 of the 5 largest QTLs detected by CIM. The independent detection of the same QTLs using two methods of analysis in separate mapping populations verifies the existence of the QTLs for BLS and provides markers to ease their introduction into elite varieties. Received: 13 October 1999 / Accepted: 29 October 1999     

FACE条件下大气O3浓度增高对小麦花后剑叶显微结构的影响

2008-2009年,应用FACE研究平台,采用烟农19、扬麦16、嘉兴002和扬辐麦2号4个小麦品种,以O3自然浓度为对照,研究了大气O3浓度增高50％对不同基因型小麦剑叶显微结构、叶绿素含量和粒重的影响.结果表明:在开花期,大气O3浓度增高对小麦产生轻微伤害,剑叶的叶肉细胞和叶绿体结构开始遭到破坏,基粒片层断裂并且开始松散;花后21 d,与对照的叶片微结构差异显著增大,剑叶内膜系统基本解体,叶绿体基本解散,基粒片层消失,加速了剑叶的衰老进程,剑叶的叶绿素含量、净光合速率和成熟期粒重均低于对照.小麦剑叶微结构和光合功能对大气O3浓度增高的响应存在明显的基因型差异,嘉兴002对O3胁迫的耐受性较好,而扬辐麦2号对O3胁迫的反应较敏感.     

Novel pleiotropic loci controlling panicle architecture across environments in japonica rice （Oryza sativa L.）

To identify quantitative trait loci (QTLs) controlling panicle architecture in japonica rice, a genetic map was constructed based on simple sequence repeat (SSR) markers and 254 recombinant inbred lines (RILs) derived from a cross between cultivars Xiushui 79 and C Bao. Seven panicle traits were investigated under three environments. Single marker analysis indicated that a total of 27 SSR markers were highly associated with panicle traits in all the three environments. Percentage of phenotypic variation explained by single locus varied from 2% to 35%. Based on the mixed linear model, a total of 40 additive QTLs for seven panicle traits were detected by composite interval mapping, explaining 1.2%-35% of phenotypic variation. Among the 9 QTLs with more than 10% of explained phenotypic variation, two QTLs were for the number of primary branches per panicle (NPB), two for panicle length (PL), two for spikelet density (SD), one for the number of secondary branches per panicle (NSB), one for secondary branch distribution density (SBD), and one for the number of spikelets per panicle (NS), respectively. qPLSD-9-1 and qPLSD-9-2 were novel pleiotropic loci, showing effects on PL and SD simultaneously. qPLSD-9-1 explained 34.7% of the phenotypic variation for PL and 25.4% of the phenotypic variation for SD, respec- tively. qPLSD-9-2 explained 34.9% and 24.4% of the phenotypic variation for PL and SD, respectively. The C Bao alleles at the both QTLs showed positive effects on PL, and the Xiushui 79 alleles at the both QTLs showed positive effects on SD. Genetic variation of panicle traits are mainly attributed to additive effects. QTL × environment interactions were not significant for additive QTLs and additive × additive QTL pairs.     

Leaf area estimation by simple measurements and evaluation of leaf area prediction models in Cabernet-Sauvignon grapevine leaves

For two growing seasons (2005 and 2006), leaves of grapevine cv. Cabernet-Sauvignon were collected at three growth stages (bunch closure, veraison, and ripeness) from 10-year-old vines grafted on 1103 Paulsen and SO4 rootstocks and subjected to three watering regimes in a commercial vineyard in central Greece. Leaf shape parameters (leaf area-LA, perimeter-Per, maximum midvein length-L, maximum width-W, and average radial-AR) were determined using an image analysis system. Leaf morphology was affected by sampling time but not by year, rootstock, or irrigation treatment. The rootstock×irrigation×sampling time interaction was significant for all the leaf shape parameters (LA, Per, L, W, and AR) and the means of the interaction were used to establish relationships between them. A highly significant linear function between L and LA could be used as a non-destructive LA prediction model for Cabernet-Sauvignon. Eleven models proposed for the non-destructive LA estimation in various grapevine cultivars were evaluated for their accuracy in predicting LA in this cultivar. For all the models, highly significant linear functions were found between calculated and measured LA. Based on r ² and the mean square deviation (MSD), the model proposed for LA estimation in cv. Cencibel [LA = 0.587(L×W)] was the most appropriate.     

OsCDPK13, a calcium-dependent protein kinase gene from rice,is induced by cold and gibberellin in rice leaf sheath

Calcium-dependent protein kinases (CDPKs) play an important role in rice signal transduction, but the precise role of each individual CDPK is still largely unknown. Recently, a full-length cDNA encoding OsCDPK13 from rice seedling was isolated. To characterize the function of OsCDPK13, its responses to various stresses and hormones were analyzed in this study. OsCDPK13 accumulated in 2-week-old leaf sheath and callus, and became phosphorylated in response to cold and gibberellin (GA). OsCDPK13 gene expression and protein accumulation were up-regulated in response to GA₃ treatment, but suppressed in response to abscisic acid and brassinolide. Antisense OsCDPK13 transgenic rice lines were shorter than the vector control lines, and the expression of OsCDPK13 was lower in dwarf mutants of rice than in wild type. Furthermore, OsCDPK13 gene expression and protein accumulation were enhanced in response to cold, but suppressed under salt and drought stresses. Sense OsCDPK13 transgenic rice lines had higher recovery rates after cold stress than vector control rice. The expression of OsCDPK13 was stronger in cold-tolerant rice varieties than in cold-sensitive ones. The results suggest that OsCDPK13 might be an important signaling component in the response of rice to GA and cold stress.     

Mapping of qGL7-2,a grain length QTL on chromosome 7 of rice

A residual heterozygous line(RHL)carrying a heterozygous segment between two SSR loci RM11 and RM134 on the rice chromosome 7 was selected from a set of recombinant inbred lines from the cross D50(javanica)/HB277(indica).The former parent produces much longer grains than the latter.Selfed progenies of this selection were analyzed genotypically(SSRs)and phenotypically(grain length).Grain length was discontinuously variable in the mapping populations,allowing for the placement of this QTL qGL7-2 within a～4.8 cM interval defined by RM351 and RM234.A set of new markers within this region were developed,which narrowed the QTL to a 278 kb region defined by the markers Indel1 and RM21945.This region contains 49 predicted genes.The results also suggest that the novel allele for grain length will be used for the application of marker assisted selection for the improvement of grain length.