拟南芥春化相关基因VRN2的克隆及其植物表达载体的构建

本文利用特异性引物,从拟南芥RNA中提取春化相关基因VRN2 cDNA序列,GenBank登录号AY063047。该基因序列大小为1 354 bp,编码区为1 323 bp,编码氨基酸441个。将克隆片段插入中间载体pBPFΩ7,经PstI酶切回收带有P35S启动子和nos终止子片段,连接pBI121载体,构建VRN2基因的植物表达载体。     

小麦品种耐寒性与春化VRN-1等位基因的关系研究

以来自11个省份的134个小麦品种为试验材料,于2009-2010年在石家庄种植,调查冻害情况;并利用分子标记鉴定VRN-1等位基因组成,以明确小麦品种春化VRN-1等位基因组成与耐寒性的关系。结果表明:小麦品种的耐寒性与其VRN-1等位基因组成、地理来源和耐旱性等因素有密切关系。当地理来源相同时,品种的耐寒性一般随着VRN-1等位基因控制的春化效应的增强而减弱;当VRN-1等位基因组成相同时,品种的耐寒性一般随着地理来源的纬度降低而减弱。本研究结果为小麦品种的耐寒性改良提供了重要参考。     

三个小麦春化基因的时空表达特性分析(英文)

为明确小麦春化基因的时空表达特性,以中国春和洛旱2号小麦品种为试验材料,利用半定量RT-PCR技术,分析了3个春化基因VERNALIZATION1(VRN1)、VRN2和VRN3的时空表达特性。结果表明,VRN1在中国春的三叶期叶片和根、灌浆期的茎秆和旗叶、花药、胚珠和发育的种子中均有不同程度的表达。在开花前,表达水平呈上升趋势,而花后呈降低的趋势,在干种子和萌发种子的胚芽中没有检测到表达;在洛旱2号中,除了在三叶期的叶片和根中没有检测到表达外,VRN1的表达特性与中国春有相同的趋势。VRN2只在三叶期的叶片和萌发种子的胚芽中表达,在其他检测的组织中没有表达;VRN3的表达与VRN1的时空表达特性相似,但在根中未检测到表达。这一结果为进一步分析普通小麦品种春化发育的分子调控机理提供了重要信息。     

黑龙江小麦春化和光周期主要基因组成分析

选取黑龙江省小麦品种126份,对其春化和光周期基因型及农艺性状进行研究。结果表明,春化和光周期基因位点显性等位变异组合在黑龙江省小麦中分布频率明显不同。含有显性基因组合Vrn-A1/Vrn-D1的分布频率最高,为26.2%,其次是显性基因Vrn-A1/Vrn-B1和Vrn-A1/Vrn-B1/Vrn-D1,分布频率分别为23.8%和23.0%,最低的是Vrn-B1基因,分布频率为0.8%,Vrn-B3位点在黑龙江小麦中不存在显性等位变异。光周期基因Ppd-D1位点的检测结果表明,53个小麦品种携带有Ppd-D1a基因型,表明光钝型小麦占42%,73个品种携带Ppd-D1b基因型,表明光敏型小麦占58%。结合田间性状调查分析春化和光周期基因对农艺性状的影响,发现在黑龙江省小麦品种中,光周期基因型对小麦的抽穗期有影响,Ppd-D1a的抽穗期比Ppd-D1b的抽穗期提前1~5d;春化和光周期基因等位变异组合对苗期习性有影响。     

河南小麦新品种的春化基因型鉴定

为了了解河南省最新培育小麦品种春化基因的等位变异状况,本研究利用分子标记技术对河南省新培育的50份冬小麦新品系(种)的春化基因Vrn-A1、Vrn-B1、Vrn-D1和Vrn-B3位点的等位变异组成进行了鉴定和分析。结果表明,所有参试小麦品种的Vrn-B3位点基因型均表现为隐性,48份小麦品种的Vrn-A1和Vrn-B1位点基因为隐性,42份小麦品种的Vrn-D1位点基因为隐性,说明隐性基因在河南小麦中占据主导地位。其中,豫农2019、豫农2020、豫农2071、国麦301、平安08-8、百农69、囤麦3698、08漯33共8个小麦品种的Vrn-D1位点基因均为显性的Vrn-D1a类型。小麦品系豫农2053和豫农3052的Vrn-A1和Vrn-B1位点的春化基因均表现为缺失,进一步研究表明,这2份小麦新品系仍能正常开花,但开花期比对照周麦18分别晚1d和2d,因此Vrn-A1和Vrn-B1并非小麦开花的必需基因。本研究将为黄淮麦区广适、高产小麦新品种的选育和推广提供参考。     

与拟南芥春化作用相关的基因及其春化记忆模型

植物能感应春化并记住这一效应,且通过一系列的信号传导,最终调控开花。文章就拟南芥的春化相关的基因,春化记忆分子模型、春化记忆与开花调控途径以及与此不同的另一种春化记忆模型和小麦春化记忆分子机制的研究进展作了介绍。     

高羊茅春化基因FaVRN1亚细胞定位与差异表达分析

高羊茅在生长季出现生殖枝,抑制新枝形成,不利于草坪质量及其持久性生长.研究春化基因的分子特征,探索抑制生殖生长的分子育种新途径,对坪用型高羊茅品种改良具有重要意义.本研究在克隆高羊茅春化基因FaVRN1的基础上,构建高羊茅春化基因FaVRN1与绿色荧光蛋白基因GFP融合的植物表达载体p-FaVRN1-hGFP,利用基因枪转化法转入洋葱表皮细胞,荧光显微镜检测融合基因的瞬时表达,并运用实时荧光定量PCR分析春化基因FaVRN1在春化与非春化条件下的表达差异.研究结果表明,FaVRN1基因编码的蛋白产物位于细胞核,符合它作为转录因子特性;春化条件下,FaVRN1基因的表达随处理时间延长逐渐增加.非春化条件下,FaVRN1基因的表达随处理时间延长而降低.FaVRN1基因在春化条件下的表达水平远高于非春化条件,FaVRN1基因的表达受春化条件正调控.     

黄淮南片冬麦区主导品种春化基因及冬春性分析

以1950～2007年黄淮南片冬麦区的127个主导小麦品种为材料,利用第5同源群的春化基因分子标记对其进行了春化基因检测,并分析了小麦品种的春化基因与其冬春性的对应关系及黄淮南片冬麦区8次品种更换中春化基因与品种冬春性的演变规律.结果表明,参试品种中没有品种携带显性Vrn-A1基因,7个品种含有Vrn-B1基因(5.5%),2个品种含有Vrn-B1+Vrn-D1基因(1.6%),56个品种含有Vrn-D1基因(44.1%).春化基因类型与品种冬春特性基本相符,春化基因控制着小麦品种的冬春特性.主导品种含春化显性基因频率的变化趋势与冬春性变化规律存在较大差异,与传统方法相比,仅用春化基因来确定品种冬春性存在一定的不完善之处.采用春化基因分子标记与传统的冬春性鉴定方法相结合来认识品种冬春性、预测品种的抗寒性对黄淮南片冬麦区的小麦品种利用更具有指导意义.     

4个春性基因型小麦品种与‘京841’杂交F_1代的表型分析

选用4个具有不同显性春化基因型的小麦品种与冬性小麦品种‘京841’进行杂交实验,通过显性春化基因特异性PCR分析技术鉴定杂交F1代植株,并分析4个杂交组合的正反交F1代植株表型特性。结果显示,各显性春化基因已经导入到各杂交F1代植株中,且其苗穗期受显性春化基因的控制而有效缩短;3个杂交组合的F1代穗粒数在正反交之间存在显著差异,推测穗粒数受细胞质遗传因素的影响较大,其中以‘新春2号’和‘豫麦18’分别为母本和父本与‘京841’杂交后F1代的穗粒数表现出较强的杂种优势,4个杂交组合的F1代千粒重均表现出较强的杂种优势。     

抗除草剂草甘膦EPSPs基因在小麦中的转化

通过基因枪法,用抗除草剂草甘膦的ＥＰＳＰｓ基因转化小麦京花１号的幼穗约１０００个,及幼胚约８００个,经草甘膦选择后分别获得３８株和４株再生植株。这些再生植株经ＰＣＲ和（或）Ｓｏｕｔｈｅｒｎ杂交证明,其中有部分再生植株的基因组中稳定整合了外源ＥＰＳＰｓ基因,并且转化体中部分是可育的。首次用实验证明,抗除草剂草甘膦的ＥＰＳＰｓ基因作为单子叶禾谷类作物小麦基因转化的选择标记是行之有效的。     

河北省小麦品种基于农艺性状的遗传多样性分析

为明确河北省小麦品种遗传多样性的基础和历史演变,本研究以所收集的该省近60年来审(认)定的125个小麦品种为研究材料,以8个农艺性状为基础进行了遗传多样性分析.结果表明:河北省小麦品种农艺性状变异比较丰富,以每穗不孕小穗数的变异系数最高(52.63%),其次为单株粒重和有效分蘖数,每穗小穗数的变异最小;多样性指数从20世纪70年代的1.62增加到目前的1.89,平均值为1.76,但略低于其他省份.在上述分析基础上,用最长距离法可将125份材料聚为3大类,其中80.8%(101个)的品种归入第3大类,主要为20世纪90年代以后的品种.这说明该省小麦品种遗传多样性在逐步提高的同时,其遗传基础仍需进一步拓宽.     

Aluminum Partitioning in Intact Roots of Aluminum-Tolerant and Aluminum-Sensitive Wheat (Triticum aestivum L.) Cultivars

Aluminum (Al) partitioning in intact roots of wheat (Triticum aestivum L.) cultivars that differ in sensitivity to Al was investigated. Roots of intact seedlings were exposed to Al for up to 24 hours and distribution of Al was assessed visually by hematoxylin staining or by direct measurement of concentration of Al by atomic absorption spectrophotometry or ion chromatography. Major differences in Al accumulation between Al-tolerant (Atlas 66) and Al-sensitive (Tam 105) cultivars were found in the growing regions 0 to 2 and 2 to 5 millimeters from the root apex. Al content was 9 to 13 times greater in the 0 to 2 millimeters root tips of cv Tam 105 than in the tips of cv Atlas 66 when exposed to 50 micromolar Al for 19 to 24 hours. The oxidative phosphorylation inhibitor carbonyl cyanide m-chlorophenylhydrazone and the protein synthesis inhibitor cycloheximide increased Al uptake by intact root tips of cv Atlas 66. Also, loss of Al from the roots of both cultivars was measured after the roots were “pulsed” with 50 micromolar Al for 2 hours and then placed in an Al-free nutrient solution for 6 hours. The 0 to 2 millimeter root tips of cv Tam 105 lost 30% of the absorbed Al, whereas the tips of cv Atlas 66 lost 60%. In light of these results, we conclude that the differential Al sensitivity in wheat correlates with the concentration of Al in the root meristems. The data support the hypothesis that part of the mechanism for Al tolerance in wheat is based on a metabolism-dependent exclusion of Al from the sensitive meristems.     

普通小麦抗倒伏相关性状的全基因组关联分析

解析小麦抗倒伏遗传机制,发掘抗倒伏位点,选育抗倒伏品种,对于保障我国粮食生产安全具有重要意义.周8425B是我国黄淮麦区重要的小麦骨干亲本,农艺性状优良,其衍生品种具有较好的抗倒伏特性.本研究对周8425B及其衍生品种等62份材料的8个抗倒伏相关性状进行评价,并结合50K SNP芯片数据进行全基因组关联分析(GWAS,...     

杂交后代中转基因小麦外源1Ax1基因的遗传

高分子量麦谷蛋白亚基1Ax1基因是决定小麦加工品质的主效基因之一,在小麦胚乳中增加1Ax1基因的表达量可以提高其加工品质,这对于小麦的品质改良具有重要意义。采用外源1Ax1基因超量表达的转基因小麦‘B102-1-2’为父本,常规小麦品种‘鄂麦12’和‘川89-107’为母本进行杂交试验。采用SDS-PAGE技术检测并分析各组合亲本、F1代、F2代的HMW-GS组成,从而研究转基因小麦‘B102-1-2’中外源品质基因1Ax1表达的遗传规律。研究结果表明:外源基因有效地整合进入主栽小麦的基因组中,并且正确表达,在F2代中表现出15∶1的分离比,遵循孟德尔遗传模式,这对于杂交育种策略的选择制订具有指导意义。     

Response of Spring Wheat (Triticum aestivum L.) Quality Traits and Yield to Sowing Date

The unpredictability and large fluctuation of the climatic conditions in rainfed regions do affect spring wheat yield and grain quality. These variations offer the opportunity for the production of better quality wheat. The effect of variable years, locations and sowing managements on wheat grain yield and quality was studied through field experiments using three genotypes, three locations for two years under rainfed conditions. The two studied years as contrasting years at three locations and sowing dates depicted variability in temperature and water stress during grain filling which resulted considerable change in grain yield and quality. Delayed sowing, years (2009–10) and location (Talagang) with high temperature and water stress resulted increased proline, and grain quality traits i.e. grain protein (GP) and grain ash (GA) than optimum conditions (during 2008–09, at Islamabad and early sowing). However, opposite trend was observed for dry gluten (DG), sodium dodecyl sulphate (SDS), SPAD content and grain yield irrespective of genotypes. The influence of variable climatic conditions was dominant in determining the quality traits and inverse relationship was observed among some quality traits and grain yield. It may be concluded that by selecting suitable locations and different sowing managements for subjecting the crop to desirable environmental conditions (temperature and water) quality traits of wheat crop could be modified.     

Free Polyamines in Senescing Wheat(Triticum aestivum L.)

The free polyamine content of flag leaves, peduncles, rachis,glumes, and grains of wheat (Triticum aestivum L., cv. Castell)plants, ripening under field conditions, has been investigatedduring three consecutive growing seasons. Putrescine was quantitativelythe most important of all polyamines detected in these organs.Concentrations were highest in the grains, glumes and flag leaves.No correlation was found between polyamine content and the onsetof senescence of flag leaves and other organs. Excised primaryleaves, however, showed a decrease in polyamine content in thedark and also in light/dark cycles, but in the latter case onlyafter an initial increase. Sink removal of otherwise intactwheat plants caused an accumulation of putrescine in flag leavesat the later stages of senescence, whereas removal of all otherleaves was without any significant effect. Putrescine was alsorecovered in phloem-exudate samples collected throughout theperiod of grain development. In both grains and glumes, peakconcentrations of polyamines were found early during seed development. Key words: Triticum aestivum, polyamines, ripening, senescence     

Agrobacterium-Mediated Gene Delivery in Various Tissues and Genotypes of Wheat (Triticum aestivum L.)

Agrobacterium-mediated wheat transformation has been attempted employing various bacterial strains, different tissues and explants, and different methods of co-cultivation. Of the various tissues/explants employed, i.e., leaf base, actively growing callus cells, mature seeds and seedlings punctured at the mesocotyl region, the mature seeds emerged as the most suited material for Agrobacterium-mediated gene delivery. Strain A281 of A. tumefaciens was found to be most effective followed by strains A348 and GV2260. Although the GUS activity in tissues/explants co-cultivated with strain GV2260 was relatively low, it indicated an efficient splicing of the intron (inserted in the coding region of the uidA gene). Various genotypes, namely, HD2329, Arjun and CPAN1676 were also screened for susceptibility against Agrobacterium strains and NPTII activity could be demonstrated in all of them, with minor variations.     

不同处理条件下小麦胚芽鞘长的遗传分析

以小麦双单倍体(DH)群体(旱选10号×鲁麦14)的150个株系及其亲本为材料,对不同处理条件下的胚芽鞘长、幼苗株高和幼苗生物学产量进行遗传分析,以探讨胚芽鞘长、幼苗株高及幼苗生物学产量的遗传基础及其相关性.结果表明:(1)随着播种深度的增加,胚芽鞘长、幼苗株高显著增加,16.1%PEG-6000处理条件下的幼苗株高显著低于对照,干旱胁迫条件下的幼苗生物学产量显著低于正常供水对照;(2)胚芽鞘长、幼苗株高、幼苗生物学产量的遗传力分别为92.5%、90.3%和78.1%;(3)胚芽鞘长、幼苗株高及幼苗生物学产量均受多基因控制,决定各性状的基因估计数目分别为7、8和14对;(4)根据对性状偏度和峰度系数的估算结果,控制这3个性状的多基因之间均有互补作用;胚芽鞘长与幼苗株高、生物学产量及成熟期株高之间呈显著正相关.     

部分耐盐小麦品种(系)SSR位点遗传多样性研究

选择有多态性的32对SSR引物对80个小麦耐盐品种(系)进行遗传差异研究,共检测出155个等位变异,平均每个位点上有4.75个等位变异;供试80份耐盐小麦品种(系)来源广泛,遗传基础丰富,表现出较高的遗传多样性,遗传相似系数范围在0.26～0.81;聚类分析结果显示,冬性小麦品种(系)聚为一大类;春性小麦品种(系)也聚为一大类;一些系谱相同或相近的品种(系)遗传相似系数较大;A、B、D基因组中SSR位点平均等位变异差异不大,以B基因组较高.