玉米杂交种与亲本雌穗花器官形成期蛋白差异表达谱分析

为探讨玉米穗粒数杂种优势形成的分子机制,以玉米强优势杂交种豫玉22及其亲本综3和87-1的花器官形成期雌穗为材料,采用高通量的2-DE和MALDI TOF MS技术,建立了它们的蛋白差异表达谱,并对差异蛋白进行了质谱鉴定。结果显示,在检测到的1 290个蛋白点中,有114个(8.84%)在杂交种与亲本之间的表达差异达到显著水平,其中表现单亲沉默、偏高亲、中亲表达、偏低亲、杂种上调、杂种下调、亲本特异和杂种特异表达模式的蛋白点分别为27、25、15、13、11、11、10和2个。另外,成功鉴定出其中的104个差异表达蛋白点,涉及代谢、信号转导、能量、转录、蛋白质合成、蛋白运输与储藏、细胞生长与分裂、细胞结构、抗病防御、次生代谢、转座子及功能未知和假定蛋白等12个功能类别。玉米杂交种与亲本蛋白在丰度上的明显差异及涉及多个功能类别,可能与玉米穗粒数杂种优势的形成有关。     

矮杆籼稻与光壳稻杂交若干性状与亲本的相关性和配合力分析

用6个矮籼与9个光壳稻杂交配组35个组合，通过分析杂种F1代杂种优势与亲本的相关性。结果表明：矮籼与光壳稻杂交F1代具有明显的优势，但组合间和性状间存在差异；杂种F1的生育期与中亲值显著正相关；有效稳与矮籼，光壳稻亲本均呈显著正相关，与中亲值也呈显著正相关；结实率与光壳亲本和中亲值均呈显著正相关。配合力分析表明，矮籼80－66，85－1210及光壳稻中水15等是杂种优势利用的良好亲本。     

基于蛋白质组学的高丹草苗期杂种优势分析

高丹草是代表性的利用杂种优势的饲用牧草, 本研究以杂种高丹草及其亲本三叶期叶片为试材, 采用双向电泳、质谱技术及生物信息学分析方法, 进行了蛋白质组学研究。凝胶上检测到的可重复蛋白点400多个, 其中杂种与亲本间达到了显著水平的差异蛋白点34个, 包括显性( 单亲沉默3个, 偏高亲表达17个, 偏低亲表达5个)和超显性表达( 特异表达1个, 超高亲表达6个, 超低亲表达2个)模式, 因此推测显性和超显性效应共同促进高丹草杂种优势的形成, 且显性效应作用更大。同时, 成功鉴定出其中的27个蛋白点涉及到8个功能类别, 即：光合作用、碳水化合物代谢、胁迫响应、ATP合成、蛋白质合成、电子转移、信号转导及未知蛋白。高丹草所占比例最大的光合蛋白多数呈上调表达, 表明杂种叶片光合作用增强进而同化更多的有机物是杂种优势形成的主要原因。网络互作的关键节点蛋白为杂种优势特异蛋白的基因操作提供了靶蛋白。本研究在蛋白质水平为高丹草杂种优势分析提供了理论依据, 也为其他饲草作物的相关研究提供了理论参考。     

鹅掌楸属植物生长旺盛期叶芽基因差异表达与杂种优势关系的分析

为了探讨鹅掌楸属植物杂种优势形成的分子机理,以其亲本及其杂种生长旺盛期的芽为材料,利用DDRT-PCR技术,分析了杂种及亲本基因表达的差异,并与杂种的4个生长性状杂种优势表现进行相关分析。结果发现,杂种和亲本之间存在显著的基因表达差异,可归纳为：双亲共沉默型（Ⅰ）、杂种特异表达型（Ⅱ）、单亲表达一致型（Ⅲ）和单亲表达沉默型（Ⅳ）。通过相关分析发现：单亲表达一致型（Ⅲ）与杂种鹅掌楸的叶面积杂种优势呈显著正相关;单亲表达沉默型（Ⅳ）与杂种鹅掌楸的地径和株高都呈显著负相关。进而推测,基因的差异表达可能与鹅掌楸属植物甚至林木杂种优势的形成相关。     

玉米雌穗发育杂种优势相关miRNA的研究

杂种优势已广泛应用于玉米育种, 在提高玉米产量、品质以及增强抗逆性等方面起到了重要的作用, 然而杂种优势的分子机制尚不清楚。植物miRNA主要在转录和转录后水平调节基因的表达。为阐明miRNA是否及如何对玉米雌穗发育杂种优势产生影响, 本研究对玉米杂交种郑单958及其亲本自交系(昌7-2和郑58)进行了高通量miRNA测序和降解组测序。取玉米雌穗花序分生组织(IM)发育为成对小穗分生组织(SPM), 进而产生小穗分生组织(SM), 及小花分生组织(FM)将3个不同时期的雌穗样品用于miRNA建库测序, 鉴定出16个miRNA家族中的81个保守miRNA为非加性表达, 认为是与雌穗发育杂种优势形成相关的miRNA; 3个阶段中分别检测到80.30%、56.06%和48.10%的非加性表达的miRNA被显性或超显性抑制。鉴定出8种新的miRNA, 属于7个miRNA家族。通过雌穗降解组测序, 发现在郑单958及其亲本自交系中共同检测到的miRNA靶向42个基因的82个转录本。根据测序结果构建了miRNA参与玉米雌穗杂种优势的模型, 并推测在雌穗发育早期阶段杂交种雌穗的miRNA的普遍抑制导致其靶基因上调表达, 随着发育进程miRNA逐步解除抑制, 带来其靶基因表达量的逐步减少, 这种miRNA与其靶基因的拮抗关系也许与玉米雌穗发育杂种优势形成有关。     

矮秆小麦亲本材料茎叶特性遗传分析

以5个矮秆小麦材料为母本,6个高产小麦为父本,采用不完全双列杂交设计,对小麦9个茎叶性状和6个穗部性状进行遗传分析。结果表明:9个茎叶性状的表现都是由加性和非加性基因共同决定的。各性状中遗传力均较高,除叶宽和结实小穗两性状外,其余性状的G.C.V均较大,表明对其选择和改良的潜力较大。大多数亲本的自身表现与一般配合力密切相关,绵阳01821、郑矮4号、绵阳04854具有很强的致矮力,是很好的致矮亲本。本文还探讨了各茎叶性状与穗部性状一般配合力的相关关系,指出小麦高产育种应注意协调好提高结实小穗、穗长、每穗粒数与提高穗下节长及穗颈节长间的关系。     

旱地杂种小麦优势测定及亲本组配模式研究

在旱作条件下,选用水旱两种生态型的小麦品种各4个,组成4×4不完全双列杂交,分折了8个主要农艺性状的超标优势和杂种优势,并对其遗传机制进行研究.结果表明水旱品种间杂交,其F1杂交优势普遍存在.F1杂种生态型是以千粒重为主,兼顾穗粒数,表现为重穗型.杂种F1产量性状变异受水地亲本影响大,旱地亲本对株高、穗下节长,单株穗数等与抗旱性密切的性状影响明显.水旱两种生态型品种杂交,易于实现抗旱与丰产的有机结合,是一种值得重视的抗旱育种杂交模式.     

中棉所12及其选系配制的4个杂交棉幼苗期基因差异表达

以与中棉所12及其2个选系为亲本组配的4个杂交棉中棉所28、中棉所29、湘杂棉2号和冀棉18苗期的根和顶端叶为研究材料,采用cDNA-AFLP技术分析苗期杂交棉与亲本的根和叶基因差异表达,并用QuantitativeReal-Time技术加以验证.结果表明:(1)在4个杂交组合中,中棉所12选系是营养生长杂种优势高值亲本;(2)杂交种和亲本间存在显著的基因表达差异,可分为杂交种上调、单亲显性、单亲沉默、杂交种下调4种表达型.4个杂交组合在三叶期根和叶中差异表达基因的4种类型比例趋势基本一致,单亲差异表达型(包括显性和沉默表达)在根和叶中所占比例较高,杂种下调表达型所占比例较低,反映出苗期单亲差异表达型在杂种优势形成中起主要作用;叶部差异表达基因数目和比例(29.20%～46.09%)比根(15.65%～22.49%)高的多,说明叶中基因差异表达可能比根中基因差异表达对杂种优势形成作用更大;(3)高值亲本中棉所12选系与杂交棉共同表达的基因多于低值亲本与杂交棉共同表达的基因,从分子水平上证明中棉所12选系在杂交棉冀棉18、中棉所29和中棉所28的苗期营养生长杂种优势产生中起优势亲本的作用;(4)4种杂交组合差异表达基因(包含叶和根)占总表达基因的27.00%～34.56%,分析差异表达基因类型和4个杂交组合的关系发现,超显性效应占3.30%～7.17%,超低亲效应占2.62%～4.14%,低亲效应占5.65%～13.03%,显性效应和加性效应是主要的杂种优势效应,占79.52%～83.79%.多种杂种优势效应的并存说明杂种优势可能是多基因共同作用产生多种效应的结果;(5)超亲优势组合中棉所28的超显性效应占7.17%,明显高于其他3个表现中亲优势组合,说明杂交种上调表达型可能对苗期杂种优势产生起重要作用.     

群体密度对提型杂种小麦产量性状及其杂种优势的影响

本文研究了群体密度对提型杂种小麦产量性状及其杂种优势的影响.结果表明:各性状在密度间都存在着显著差异.单株穗数、穗粒数和千粒重随密度增加而下降,籽粒产量与密度呈二次抛物线关系.平均穗粒数和单株穗数的杂种优势在产量杂种优势形成中的直接作用是低密度高于高密度;千粒重的杂种优势在产量杂种优势形成中的直接作用是低密度低于高密度;通过对杂种小麦产量及杂种优势标准化联立方程组求解和对产量回归方程求导的方法,确定了杂种小麦的适宜密度范围为每亩8.5～13万株,最后提出了适当降低杂种小麦密度应采取的栽培措施.     

矮腥黑粉菌胁迫的小麦全基因组重测序与转录组联合分析

为揭示矮腥黑粉菌胁迫下的小麦抗感品种间遗传背景差异,筛选小麦抗矮腥黑穗病候选基因。本研究以小麦感病品种‘伊犁053’和中抗品种‘中麦175’为研究对象,在接种矮腥黑粉菌后对籽粒组织进行重测序与转录组测序的联合分析。经过差异InDel分析后,共有69 840个变异基因与转录组数据进行了联合分析,有7 300个基因在小麦发病籽粒组织中有表达,有162个(2.22%)基因呈极显著差异表达,且基因表达量倍数变化≥1.5倍(‘中麦175’相对于‘伊犁053’,有125个上调, 37个下调);经过差异SNP分析后,共有98 331个变异基因与转录组数据进行了联合分析,有7 438个基因在小麦发病籽粒组织中有表达,有126个(1.69%)基因呈极显著差异表达,且基因表达量倍数变化≥1.5倍(‘中麦175’相对于‘伊犁053’,有86个上调, 40个下调)。本研究成功揭示了在矮腥黑穗病胁迫下抗感病小麦品种间的遗传差异表达信息,为进一步筛选小麦抗矮腥黑穗病候选基因提供参考。     

Differential gene expression between wheat hybrids and their parental inbreds in seedling leaves

Differential display of mRNA was used to analyze the differences of gene expression in seedling leaves between heterotic hybrid/nonheterotic hybrid and their parental inbreds in order to study the molecular basis of heterosis in wheat. The results indicated that patterns of gene expression in hybrids differ significantly from their parents. Both quantitative and qualitative differences were observed. The quantitative differences include gene over-expression, gene under-expression in hybrid and dominant expression of highly-expressed parental genes in hybrids. The qualitative differences include silencing in hybrids of genes expressed either in male or female parent, and silencing in hybrids of genes expressed in both parents. Expression in hybrid of genes only expressed either in male or female parent was also observed. It was also found that some genes expressed at high level in heterotic hybrid were underexpressed or expressed at low level in nonheterotic hybrid. One differentially expressed cDNA fragment 4B was cloned and sequenced after being confirmed through Northern blot analysis. Homology search in GenBank proved that the cDNA fragment is a new sequence. The selection of primers for differential RNA display in wheat and the relationship between wheat heterosis and alteration of gene expression in hybrids as compared to their parental inbreds were also discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

不同优势小麦正反杂交种子与亲本自交种子发育前期基因表达差异

为探讨小麦杂种优势形成的分子机理，本研究选用优势不同的3个杂交组合,以授粉后2、6和12 d的正反杂交种子与亲本自交种子为材料，采用mRNA差异显示技术分析了不同优势正反杂交种子与亲本自交种子之间发育前期的基因表达差异。结果表明，所有三个时期正反杂交种子与亲本自交种子之间存在明显的基因表达差异，差异可归纳为特异     

Heterosis for yield and related characters in pea

Summary To determine the levels of heterosis in F1 hybrids, four current pea (Pisum sativum L.) cultivars from southern Australia were used as female parents and crossed with 18 introduced genotypes. The 22 parents, 72 F1 hybrids and, depending on the environment, either 54 or all 72 F2 families were grown in replicated plots in four environments. Grain yield, total dry matter, harvest index, branches per plant, pods per plant, seeds per pod, hundred seed weight, plant height, onset of flowering and flowering periods were evaluated. For both the F1 and F2 generation, heterosis was determined as the superiority over the mid-parent and also over the better parent. In addition, the superiority over the best commercial cultivar was calculated. Most hybrids were higher yielding than their mid-parent but were less stable in yield across environments. Four F1 hybrids were significantly higher yielding than the best parent, by up to 26%. There were significant correlations between F1 hybrid and mid-parent value for plant height, pods per plant and hundred seed weight but not for yield. Overall, grain yield heterosis was mainly due to more pods per plant in the hybrids. The level of heterosis for yield in a poor yielding environment was higher than that in a high yielding one. Both additive and non-additive gene effects were important in the expression of all studied traits. The average level of heterosis for grain yield and total dry matter in the F2 population was half of that in F1 hybrids. The low level of inbreeding depression from the F1 to the F2 generation suggested that epistatic gene action also contributed to the expression of grain yield. Some F2 populations maintained the high yield levels of the corresponding F1 hybrids.     

Heterosis in yield, endotoxin expression and some physiological parameters in Bt transgenic cotton

Although heterosis in cotton has been studied for many decades, very little is known about the performance of hybrids derived from Bt transgenic cotton parents. In order to known better the heterosis performance, yield and endotoxin expression in 20 hybrids (F₁) and their Bt transgenic parents were examined from 2002 to 2003 (Experiment 1), and the dynamics of source, sink and their ratios in a well‐performing hybrid H01 were investigated in 2004 and 2005 (Experiment 2). Results in Experiment 1 showed an average mid‐parent heterosis of 21.3% and an over check heterosis of 7.6% in lint yield. Considerable heterosis was also detected in boll numbers, boll size and Bt protein content. Of the 20 hybrids, H01 (K0215 × K643) exhibited the greatest heterosis in yield and Bt protein content in 2002 and 2003, while lint yields of H01 were increased 12.6% and 9.1% in 2004, and 11.7% and 8.9% in 2005, compared with K0215 and K643 in Experiment 2, respectively. Significant heterosis for dry matter accumulation and dry matter allocation to reproductive organs and ratio of fruiting forms/total plant (w/w) were also detected in H01. Sources (leaf area, leaf area index, leaf dry weight per plant and diurnal performance of photosynthesis), sinks (number of fruiting nodes, fruiting forms and dry weight of fruiting forms per plant) and the flow from source to sink were significantly enhanced in H01 relative to its parents. Both total N and Bt protein in H01 were higher than those in its parents. Significant correlation was also found between total N and Bt protein in the main‐stem leaves (R² = 0.877**). It is concluded that there existed considerable heterosis in yield, yield components and endotoxin expression in some Bt transgenic hybrids. Yield advantage of hybrid cotton (F₁) over parents can be attributed to improved source, sink and flow, while the enforced expression of Bt genes in hybrid cotton appeared to be due to the enhanced nitrogen level in plants.     

K,V型杂种小麦优势的研究

利用５（Ａ）×２（Ｒ）两组亲本杂交的１０个杂种及其相应的亲本材料，探讨高产条件下Ｋ、Ｖ型小麦的杂种优势。结果表明：单株产量有很强的杂种优势，７个组合超双亲平均值为７．１１％～２５．１％，其中３个为１５．７％～２５．１％，４个组合超标优势２．４％～１８．１％。Ｋ、Ｖ型杂种优势的主要表现是千粒重的增高，其次是穗粒数，有３个组合超双亲平均值。单株穗数的减少对杂种产量有影响，选择多穗亲本，对提高杂种的穗数有效     

Heterosis and combining ability for grain yield and other agronomic traits in winter triticale

Hybrid breeding is a widely discussed alternative for triticale. Heterosis as well as general (GCA) and specific combining ability (SCA) effects were estimated for eight agronomic traits. The experiment comprised 24 F₁ hybrids, produced by a chemical hybridizing agent, together with their six female and four male parents, grown in drilled plots in two locations. In comparison with the mid‐parent values, hybrids averaged a 6.4 dt/ha (10.1%) higher grain yield, 8.4% more kernels per spike, a 6.8% higher 1000‐kernel weight, 9.7% lower falling number (FN) and 4.4% greater plant height. SCA effects for grain yield were significant and ranged from 4.5 to 6.9 dt/ha for grain yield. Together with GCA x location interactions, they explained most of the variation. For 1000‐kernel weight, GCA effects were predominant. SCA and interactions with location accounted for most of the variation in FN, whereas interactions were negligible for plant height. Correlations between mid‐parent and hybrid performance and between GCA and per se performance of parents were tight for all traits except grain yield, which allows for pre‐selection of parental lines. Although the amount of heterosis in triticale at present is closer to wheat than to rye, by selecting parents for combining ability and identifying heterotic patterns, grain yield heterosis of up to 20% appears sufficiently encouraging to embark on hybrid breeding.     

Hybrid performance and heterosis in spring bread wheat, and their relations to SSR-based genetic distances and coefficients of parentage

Development of hybrids is considered to be a promising avenue to enhance the yield potential of crops. We investigated (i) the amount of heterosis observed in hybrid progeny, (ii) relative importance of general (GCA) versus specific (SCA) combining ability, and (iii) the relationship between heterosis and genetic distance measures in four agronomic traits of spring bread wheat. Eight male and 14 female lines, as well as 112 hybrids produced in a factorial design were grown in replicated trials at two environments in Mexico. Principal coordinate analysis based on Rogers' distance (RD) estimates calculated from 113 SSRs revealed three different groups of parents. Mid-parent heterosis (MPH) for grain yield averaged 0.02 t ha⁻¹ (0.5%) and varied from −15.33% to 14.13%. MPH and hybrid performance (F1P) were higher for intra-group hybrids than for inter-group hybrids, with low values observed in inter-group crosses involving two non-adapted Chinese parents. Combined analyses of variance revealed significant differences among parents and among hybrids. Estimates of GCA variances were more important than SCA variances for all traits. Tight correlations of GCA with line per se performance, and mid-parent value with F1P were observed for all traits. In contrast, correlations of MPH with RD and coefficient of parentage were not significant. It was concluded that the level of heterosis in spring wheat was too low to warrant a commercial exploitation in hybrids. SSRs proved to be a powerful tool for the identification of divergent groups in advanced wheat breeding materials.     

Relationship between hybrid performance and genetic diversity based on RAPD markers in wheat, Triticum aestivum L.

Random amplified polymorphic DNA (RAPD) markers were generated from 20 wheat, Triticum aestivum lines. Fifty-four fragments generated by six primers of a 10-mer arbitrary sequence were used to study their potential power in differentiating parents with different characteristics and predicting the yield performance of hybrids produced from these parents. Experimental results showed that the 20 wheat lines were divided into four groups. Group I was characterized by more grains per spike, group II by heavy grains and group III by more spikes per unit area and short plants; group IV was similar to group III but had a much higher biomass yield and grain yield. Hybrids from parents in different groups were generally superior to most hybrids from parents in the same group. Both yield performance and heterosis of hybrids from parents between group I and group III were much better than those of other intergroup hybrids. These results suggest that, based on RAPD markers, it is possible to differentiate wheat lines with different performances and that the classification of parents from these markers is of predictive value for developing superior hybrids. However, genetic distance (GD) based on RAPD markers was not significantly correlated with hybrid performance and heterosis. It appears to be impossible to predict hybrid performance from GD itself.     

Retrotransposon-related genetic distance and hybrid performance in sunflower (Helianthus annuus L.)

Heterosis is a main force leading the development of the hybrid seed industry in sunflower. The purpose of this study is to evaluate if heterosis effects for morphological traits among sunflower hybrids can be related to differences in the repetitive component of the genome of parental lines. The assumption is that, at least for certain traits, heterosis results from mutations in the cis-regulatory elements of genes, largely related to retrotransposon insertions and/or removals. Our experimental approach consists of a correlation study between hybrid performance and retrotransposon-related genetic distances between inbreds. Six sunflower inbred lines of different origin were crossed in a half diallel fashion; comparing parental lines and hybrids, mid parent heterosis of F₁ hybrids was evaluated for six traits. We estimated the parental genetic distances between the six inbreds on data gathered by the inter-retrotransposon-amplified-polymorphism (IRAP) protocol. Different retrotransposons previously isolated in sunflower were targeted by 11 primer pairs designed on conserved LTR domains. As a control, genetic distances were also calculated using 86 genic SNPs. We analysed the correlation between the mid-parent heterosis for each of the six traits analysed and the genetic distance (calculated on data obtained by SNP or IRAP analyses) between the parental lines. Differences between parents showed to be largely related to variations in the retrotransposon component of the genome. Retrotransposon-related genetic distance between parents resulted to be larger than that related to genic SNPs, and significantly correlated to seed yield and, at a lesser extent, to plant height and stem diameter in hybrids. The hypothesis that variations in the repetitive component of the genome, especially LTR-retrotransposons, affect the displaying of heterosis is discussed.