A-3中抗条锈新基因YrTp1和YrTp2的分子标记定位分析

【目的】半个多世纪的中国小麦育种史基本是育种家与条锈病的赛跑史。因此，筛选、鉴定、储备和利用新抗源是我国育种和资源研究中的一个长远战略性课题。【方法】利用小麦条锈菌条中31、32号生理小种，对来自小麦与十倍体长穗偃麦草［Thinopyrum ponticum (Host) Liu & Wang］的杂交后代材料A-3进行抗性遗传分析。用荧光SSR分子标记技术，鉴定所携带抗条锈病基因是否为新基因，并对其进行染色体定位研究。【结果】遗传分析表明，A-3对条中31号和32号的抗性由一显一隐2对基因控制。经过对196对微卫星引物的筛选，发现2B染色体短臂上的WMC477-167bp与显性基因紧密连锁，遗传距离为0.4 cM,将该显性基因定位于2BS上；7B染色体短臂上的WMC364-208bp与隐性基因连锁，遗传距离为5.8 cM。图位比较、系谱分析和抗谱分析表明，A-3所含抗条锈基因不同于已知抗条锈基因，暂定名为YrTp1和YrTp2。【结论】可利用A-3中与条锈病抗性紧密连锁的分子标记YrTp1和YrTp2将抗性基因转移到主栽品种中，在小麦育种和生产上发挥作用。     

第7同源染色体组对小麦气体交换及荧光参数的影响

[目的]研究7号同源染色体（7A、7B、7D）对小麦光合作用的影响,为今后采用遗传和生理生化手段进行小麦的高光效遗传改良提供一定理论依据。[方法]以山西农科院温室大棚中种植的小麦中国春及其7A、7B、7D染色体缺失植株N7A、N7B、N7D（美国堪萨斯州立大学B.S.Gill教授提供）为材料,于灌浆初期进行相关指标测定,研究小麦7A、7B、7D染色体对小麦光合特性的影响。[结果]N7A、N7B在灌浆初期的光合速率、气孔导度、原初光化学效率（Fv/Fm）、PSⅡ的实际光化学效率（ФPSⅡ）、PSⅡ的表观电子传递速率（ETR）都明显低于中国春,且差异达显著水平（P〈0.05）;N7D的光合速率、气孔导度都明显高于普通小麦中国春,且差异达显著水平（P〈0.05）;Fv/Fm、ФPSⅡ、ETR也高于中国春,但差异不显著。[结论]7A、7B染色体对光合速率有正效应,与气孔导度和光化学反应（荧光参数）均有关系;7D染色体对光合速率具有负效应,而这种负效应主要与气孔导度有关,与光化学反应（荧光参数）无关。     

四川和黄淮生态区小麦品种(系)主要产量性状比较

采用来自四川和河南两地区的主栽小麦(Triticum aestivum Linn.)品种和2012年两地部分区试小麦品系共27个材料,在河南辉县地区种植,随机区组法2次重复,生长期间和收获后调查产量结构性状,研究南方和北方生态区小麦品种产量构成的差异.比较分析结果表明:①四川品种在北方种植,其产量明显低于北方品种;②四川品种的穗粒数高于北方品种,但穗数和粒重低于北方品种;③北方的多穗型品种在北方生态区种植易获得高产,大穗型品种在产量结构合理的条件下也能获得较高产量;④单位面积穗数是北方冬麦区构成小麦产量的第一因素;⑤大穗型品种播量对单位面积穗数调节效果不明显.上述结果说明,不同类型生态区小麦最佳产量结构差异较大,北方品种高产主导因素是单位面积穗数,南方品种优势倾向于穗粒数.     

5种酚酸类物质对小麦幼苗的化感作用研究

为了研究5种酚酸类物质对小麦幼苗生长和生理指标的影响,测定了不同质量浓度的阿魏酸、对香豆酸、丁香酸、对羟基苯甲酸、香草酸5种酚酸类物质处理后小麦幼苗的苗高、根长、CAT活性、POD活性等形态、生理指标,为小麦与药用植物轮作的合理性提供依据。结果表明,小麦幼苗的可溶性糖含量均有所升高,50.00 mg/L对香豆酸和0.01、0.10、1.00 mg/L对羟基苯甲酸处理组分别比空白对照高18.79%、42.60%、42.69%、26.57%,均达显著水平;阿魏酸、对香豆酸和香草酸降低了小麦幼苗的苗高和主根长,对羟基苯甲酸增加了幼苗的主根长,但均不显著;对香豆酸处理组幼苗的可溶性蛋白含量、CAT活性、POD活性以及SOD活性均有所降低,而丁香酸处理组幼苗的可溶性蛋白含量、CAT活性、POD活性均有所升高,0.01、0.10、10.00、100.00 mg/L丁香酸处理组幼苗的可溶性蛋白含量分别比对照(蒸馏水)显著增加9.63%、11.22%、11.50%、8.13%。香草酸处理组幼苗的可溶性蛋白含量、CAT活性、SOD活性也有所增加。从研究结果可以看出,阿魏酸、对香豆酸对小麦幼苗有一定的化感抑制作用,丁香酸、对羟基苯甲酸和香草酸有一定的促进作用,但它们的化感指数均不大。     

小麦胚乳14-3-3基因的克隆及其重组蛋白的原核表达

【目的】克隆小麦籽粒胚乳14-3-3基因,并进行体外表达,为进一步研究其对籽粒生长发育的调控作用奠定基础。【方法】根据已有同源基因保守序列,设计插入限制性酶切位点的特异性扩增引物,采用RT-PCR方法扩增发育的小麦胚乳14-3-3基因,克隆测序后转入表达载体,在大肠杆菌中进行表达,并进行纯化。【结果】从开花后灌浆13-15 d的小麦品种济麦22籽粒胚乳中克隆到1个14-3-3基因,序列分析表明为非ε型,含1个777 bp的开放阅读框,编码蛋白259 aa,分子量约29 kD。核苷酸序列分析表明与小麦、水稻、玉米、大麦、大豆等主要农作物和模式植物拟南芥的14-3-3基因有较高的同源性,最高达98%,编码蛋白氨基酸长度也一致（260 aa左右）;在大肠杆菌中高效表达的重组蛋白约为30 kD,分子量大小与根据核苷酸序列推导的编码蛋白一致。从基因序列的同源性、编码蛋白的氨基酸长度、表达蛋白的分子量大小分析都说明克隆到的基因为14-3-3基因,并准确插入表达载体,得到了高效正确表达。将克隆的基因插入pET29c载体,热激转化大肠杆菌BL21-CodonPlus(DE3)-RP,得到了高效表达,但主要以包涵体形式（80%）存在。对重组蛋白进行了纯化,可溶性重组蛋白利用S-蛋白琼脂糖树脂得到纯化的蛋白,包涵体重组蛋白经变性溶解、复性后,也利用S-蛋白琼脂糖树脂得到了高度纯化的重组蛋白。【结论】利用RT-PCR技术从发育的小麦胚乳中克隆到1个14-3-3基因,并在大肠杆菌中得到了高效表达,重组蛋白经过纯化得到了纯度较高的活性蛋白。     

河北省小麦品种高分子量谷蛋白亚基组成分析

高分子量麦谷蛋白(HMW-GS)亚基组成是决定小麦加工品质的重要蛋白组分,对小麦品质具有重要影响。本研究利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)方法分析了148份河北省小麦品种的高分子量麦谷蛋白亚基(HMW-GS)组成及其历史变化。结果表明:在148份小麦品种中共检测到16种HMW-GS亚基和43种亚基组合。在Glu-A1位点,有3种亚基类型,以null和1亚基为主;Glu-B1位点有7种亚基类型,以"7+9"和"7+8"亚基为主;Glu-D1位点有6种亚基类型,以"2+12"亚基类型为主。在43种亚基组合中,以"null、7+9、2+12"组合为主,其他亚基组合尤其是与品质有关的亚基组合相对较少。从河北省小麦品种HMW-GS的历史演变来看,优质蛋白亚基及其组合虽然近年来有所提高,但仍需进一步加强。     

分子标记辅助选育小麦抗白粉病、优质高分子量麦谷蛋白亚基聚合体

现代小麦育种的目标是选育丰产、综合抗逆性好的优质小麦新品种,将分子标记技术与传统育种方法相结合可以大大提高育种效率.本研究利用与小麦抗白粉病基因Pm21紧密连锁的共显性PCR标记、以及优质高分子量麦谷蛋白亚基1Dx5 1Dy10特异的PCR标记对以小麦品种安农94212、安农92484为优质亲本和以抗白粉病小麦簇毛麦易位系为抗病亲本的杂交高代材料进行标记位点的检测,结合田间抗病性鉴定结果,筛选、培育出聚合有Pm21和1Dx5 1Dy10的抗白粉病小麦聚合体,为小麦抗病、优质育种提供了具有重要利用价值的中间材料.     

Late nitrogen fertilization affects carbohydrate mobilization in wheat

An experiment was carried out to determine how the late application of nitrogen (N) fertilizer affects the use of pre‐anthesis carbon reserves during the grain‐filling period of pot‐grown wheat with no water shortage. Increasing doses (equivalent to 0, 140, and 180 kg N ha^–1) of N fertilizer were applied, either in two amendments (stages GS20 and GS30) or in three amendments (stages GS20, GS30, and GS37, according to Zadoks scale). The management of fertilizer by combining an increased N rate with late N application was able to stimulate canopy development, to raise photosynthetic capacity and carbohydrate accumulation during the vegetative stages, and to increase grain yield. Based on the dynamics of carbohydrate accumulation in the ear, three phases were differentiated during the grain‐filling period, whose temporary pattern remained stable regardless of the fertilizer management. The net remobilization of carbohydrates started 12 d after anthesis from the leaves and 28 d after anthesis from roots and stems. The increase of the N dose with late N application allowed on one hand a lower use of the pre‐anthesis carbon reserves in favor of greater de novo photosynthesis during the grain‐filling period, and on the other hand greater relative contribution of the leaf and ear C reserves to remobilization towards the grain. Further splitting the dose increased only the relative contribution of ear C reserves. The stem contribution seemed to be independent of N applied whereas the root contribution tended to diminish with late N application.     

DROUGHT STRESS: Soil Water Availability Alters the Inter‐ and Intra‐Cultivar Competition of Three Spring Wheat Cultivars Bred in Different Eras

Competition for water generates a classic aspect of the tragedy of the commons, the ‘race for fish’, where crops must allocate more resource to acquisition of the limiting resource than is optimal for crop yield allocation. A pot experiment using a simple additive (target–neighbour) design was conducted to examine the above‐ground and below‐ground growth of three spring wheat (Triticum aestivum L.) cultivars when grown alone and in mixtures at three levels of water availability. The effects of competition and water availability were compared by observing patterns of growth, biomass allocation and below‐ground outcomes. Competitive interactions were investigated among cultivars ‘HST’, ‘GY602’ and ‘LC8275’, target plant of each cultivar grown without neighbouring plants are referred to herein as control plant and one target plant of each cultivar sown surrounded either by same or another cultivar as intra‐ or inter‐cultivar competition. Competitive ability was assessed as the response ratio (lnRR) between the target plant surrounded by six other plants and the target plant in isolation. Our results showed that the cultivar ‘HST’, released over a century ago, produced a higher biomass and grain yield than the more recently released cultivars ‘LC8275’ and ‘GY602’ when grown as isolated plants with sufficient water supply. However, competition for resources from neighbours led to target plant biomass and grain yield being significantly reduced relative to controls in all three cultivars, particularly in ‘HST’. When subjected to intra‐cultivar competition, the two recently released cultivars ‘LC8275’ and ‘GY602’ had higher grain yields and water use efficiency for grain than ‘HST’ in all three water regimes. The landrace ‘HST’ had better and significantly linear relationships between biomass and biomass allocation, root length and specific root length, whereas the recent and modern cultivars had much more water‐related species‐specific changes in root morphology and allocation patterns. These results suggest that crop traits that influence competitive ability, such as biomass allocation to roots and root plasticity in response to drought have changed in modern wheat cultivars because of breeding and selection.     

Screening and Analysis of Proteins Interacting with TaPDK from Physiological Male Sterility Induced by CHA in Wheat

To further research the regulatory network of pyruvate dehydrogenase kinase (designated as TaPDK) in physiological male-sterility (PHYMS) of wheat induced by chemical hybridizing agent (CHA) SQ-1, an anther cDNA library was constructed, and the proteins interacting with TaPDK were screened via yeast two-hybrid technique. Subsequently, a few candidate proteins in nucleotide expression levels were detected by real-time quantitative PCR. Yeast-two hybrid screening was performed by mating yeast strain Y2HGold containing BD-TaPDK bait plasmid with yeast strain Y187 including anther cDNA library plasmid. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Ade/-His/-Leu/-Trp) (QDO), and further were incubated on QDO medium containing AbA and X-α-Gal. The interactions between TaPDK and the proteins obtained from positive colonies were further confirmed by co-transformation validation. After plasmids DNA were extracted from blue colonies and sequenced, the sequences results were analyzed by bioinformatic methods. Finally, 24 colonies were obtained, including eight genes, namely non-specific lipid-transfer protein precursor (TanLTP), polyubiquitin (TaPUbi), glyceraldehyde-3-phosphate dehydrogenase, proliferating cell nuclear antigen (TaPCNA), CBS domain containing protein (TaCBS), actin, guanine nucleotide-binding protein beta subunit, chalcone synthase, and three new genes with unknown function. The results of quantitative RT-PCR showed that the expression levels of TanLTP, TaPUbi, and TaPCNA were obviously up-regulated in PHYMS anther, and TaCBS expression was only increased at the tricellular stage in PHYMS anther compared with in fertile lines. Whereas, the expression of TaPDK was obviously down-regulated in PHYMS lines. Collectively, these datas indicated that the majority of candidate proteins might be related to pollen abortion in PHYMS lines, which further suggested that TaPDK plays multiple roles in pollen development, besides participating in regulating pyruvate dehydrogenase complex activity.