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。     

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.     

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。     

Artificial neural networking to estimate the leaf area for invasive plant Wedelia trilobata

Leaf area are very important parameter for the understanding of growth and physiological responses of invasive plant species under different environmental factors. This study was conducted to build non-destructive leaf area model of Wedelia trilobata that were grown in greenhouse. Regression analysis and artificial neural network (ANN) approaches were used for the development of leaf area model with the help of leaf length and width of 262 plants samples. In selection of best method under both techniques, the lower value of mean absolute error (MAE), root mean square error (RMSE), mean absolute percentage error (MAPE) and higher value of R² were considered. According to the results it was found that ANN have higher value of (R² = 0.96) and lower value of error (MAE = 0.023, RMSE = 0.379, MAPE = 0.001) than regression analysis (R² = 0.94, MAE = 0.111, RMSE = 1.798, MAPE = 0.0005). It was concluded that error between predicted and actual value was less under ANN. Therefore, ANN model approach can be used as an alternating method for the estimation of leaf area. Through estimation of leaf area, invasive plant growth can predict under different environment conditions.     

Distinct controls of leaf widening and elongation by light and evaporative demand in maize

Leaf expansion depends on both carbon and water availabilities. In cereals, most of experimental effort has focused on leaf elongation, with essentially hydraulic effects. We have tested if evaporative demand and light could have distinct effects on leaf elongation and widening, and if short‐term effects could translate into final leaf dimensions. For that, we have monitored leaf widening and elongation in a field experiment with temporary shading, and in a platform experiment with 15 min temporal resolution and contrasting evaporative demands. Leaf widening showed a strong (positive) sensitivity to whole‐plant intercepted light and no response to evaporative demand. Leaf elongation was (negatively) sensitive to evaporative demand, without effect of intercepted light per se. We have successfully tested resulting equations to predict leaf length and width in an external dataset of 15 field and six platform experiments. These effects also applied to a panel of 251 maize hybrids. Leaf length and width presented quantitative trait loci (QTLs) whose allelic effects largely differed between both dimensions but were consistent in the field and the platform, with high QTL × Environment interaction. It is therefore worthwhile to identify the genetic and environmental controls of leaf width and leaf length for prediction of plant leaf area.     

生育前期遮光对水稻灌浆期剑叶生理特性及籽粒生长的影响

通过盆栽试验研究了生育前期（插秧至孕穗期）遮光对水稻分蘖、灌浆期剑叶生理特性、籽粒生长、产量及其构成因素的影响.结果表明：前期遮光使水稻分蘖数明显降低（降低26.72%）,齐穗期与成熟期发生延迟;遮光结束后,水稻灌浆期剑叶叶面积、可溶性糖含量分别比对照增加了33.86%和30.23%;每穗实粒数、千粒重、籽粒终极生长量、最大灌浆速率、平均灌浆速率分别比对照降低了8.65%、4.81%、9.74%、20.22%和19.13%;有效穗数与产量分别降低了25.26%和39.56%,差异显著;灌浆速率峰值的出现时间提前了1.66 d,实际灌浆时间延长了6.80 d.与弱耐荫水稻品种相比,耐荫品种灌浆前中期的剑叶叶绿素a、叶绿素b、叶绿素(a+b)含量增加,灌浆期的剑叶叶面积增加幅度大,蛋白氮、可溶性糖含量及叶绿素 a/b升高,籽粒终极生长量及千粒重接近对照,减产幅度相对较低.     

黄淮海平原冬小麦（Triticum aestivum）旗叶的生理生态特性

分别于2005年和2006年4、5月份,在中国科学院封丘农业生态试验站用Li-6400光合测定仪对田间冬小麦(Triticumaestivum)进行了净光合速率(Pn)、蒸腾速率(Tr)日变化的测定,并同步测定了有效辐射(PAR)、气温(Ta)、叶温(Tl)、空气相对湿度(Rh)、气孔导度(Gs)、胞间CO2浓度(Ci)等因子,研究了冬小麦旗叶的Pn、Tr与环境和生理因子之间的关系。结果表明,冬小麦旗叶的Pn随PAR的增加而升高,其光补偿点和光饱和点分别在50~60μmo.lm-.2s-1和1050~1100μmol.m-.2s-1左右;Tr随PAR呈线性升高。Ta与Tl的相关系数r=0.886**,达显著水平,冬小麦旗叶Pn随Ta、Tl的增加而升高,当Ta、Tl分别超过30℃、32℃左右时,Pn随两因素的增加反而下降;Tr随Ta、Tl的增加呈直线升高。冬小麦旗叶Pn、Tr与Gs的相关系数分别为r=0.407**和r=0.322**,二者均随Gs的增加而升高,当Gs超过0.2mo.lm-.2s-1左右时,Pn的速度就会减缓。冬小麦旗叶的Pn、Tr均随Ci的增加而下降。通过对生态、生理因子的多元回归分析表明,影响黄淮海平原冬小麦旗叶的Pn、Tr的主要因子是PAR、Gs,除此以外,Ci也是影响Pn的重要因子。     

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.     

水稻剑叶突变体的表型及T-DNA旁邻基因分析

从已构建的水稻（Oryza sativa L.）T-DNA插入突变体中鉴定获得一株穗部额外发育出叶片的突变体,并根据该叶片的形态学位置将其命名为剑叶突变体（J4）。研究表明这种额外发育的叶片呈现明显的缺陷,主要表现为叶片短小、表皮细胞变小、叶片中维管束数目减少等。进一步通过TAIL-PCR和inverse-PCR的方法克隆该突变体中T-DNA插入位置的旁邻序列,从而准确地将T-DNA定位到2号染色体上。基因表达分析显示,T-DNA插入位置附近的AK100376基因在J4突变体以及表型类似突变体neck leaf 1中的表达均被明显下调,可初步将其确定为与剑叶突变体表型相关的候选基因。     

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.     

Non-destructively predicting leaf area,leaf mass and specific leaf area based on a linear mixed-effect model for broadleaf species

Based on a linear mixed-effect model, we propose here a non-destructive, rapid and reliable way for estimating leaf area, leaf mass and specific leaf area (SLA) at leaf scale for broadleaf species. For the construction of the model, the product of leaf length by width (LW) was the optimum variable to predict the leaf area of five deciduous broadleaf species in northeast China. In contrast, for species with leaf thickness (T) lower than 0.10 mm, the surface metric of a leaf (e.g., LW or width) was more suitable for predicting leaf mass; and for species with leaf thickness larger than 0.10 mm, the volume metric of a leaf (e.g., the product of length, width and thickness together, LWT) was a better predictor. The linear mixed-effect model was reasonable and accurate in predicting the leaf area and leaf mass of leaves in different seasons and positions within the canopy. The mean MAE% (mean absolute error percent) values were 6.9% (with a scope of 4.1–13.0%) for leaf area and 13.8% (9.9–20.7%) for leaf mass for the five broadleaf species. Furthermore, these models can also be used to effectively estimate SLA at leaf scale, with a mean MAE% value of 11.9% (8.2–14.1%) for the five broadleaf species. We also propose that for the SLA estimation of the five broadleaf species examined, the optimum number of sample leaves necessary for good accuracy and reasonable error was 40–60. The use of the provided method would enable researchers or managers to rapidly and effectively detect the seasonal dynamic of leaf traits (e.g., leaf area, leaf mass or SLA) of the same sample leaves in the future.     

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.