小麦耐甘露醇变异系生理特性分析

目的对二次离体筛选获得的小麦耐甘露醇变异细胞系进行生理生化及分子生物学鉴定。方法检测该变异细胞系在甘露醇,NaCl和聚乙二醇(PEG-6000)模拟的渗透胁迫环境中的耐受生长能力,并测定变异系在甘露醇胁迫下游离脯氨酸含量、可溶性糖含量、可溶性蛋白含量、Na /K 含量等生理生化特性及可溶性蛋白质组成和基因组DNA多态性等分子特征。结果通过愈伤组织在甘露醇,NaCl和聚乙二醇(PEG-6000)模拟的渗透胁迫条件下的生长实验,发现变异系细胞系可以在对照细胞系不能生长的20%甘露醇,1.5%NaCl和20%PEG-6000胁迫条件下,分别表现出14.5%(见图1),12.8%(见图2),41.8%(见图3)的相对生长量;在20%甘露醇胁迫条件下变异系细胞系游离脯氨酸积累量为对照系的80%(见图4),可溶性糖积累量为对照系的1.2倍(见图5),可溶性蛋白含量为对照系的1.3倍(见表1)。在相同浓度的甘露醇模拟的渗透胁迫环境中变异系再生植株比对照植株相能维持较高的K /Na 比值(见表2)。与对照相比,耐甘露醇变异细胞系再生植株可溶性蛋白SDS-PAGE发生显著变化:6条新可溶性蛋白谱带出现在变异系再生植株中,同时对照系中的1条可溶性蛋白谱带在突变株中缺失(见图6)。变异系植株与对照株RAPD带型呈现一定的多态性(见图7),表明变异系基因组DNA与对照相比发生了突变。结论所研究的小麦耐甘露醇变异细胞系是一个具有较强渗透胁迫耐受能力,可用于进一步育种工作的良好突变体材料。     

Genetic diversity and construction of core collection in Chinese wheat genetic resources

Genetic diversity among 5029 accessions representing a proposed Chinese wheat core collection was analyzed using 78 pairs of fluorescent microsatellite （SSR） primers mapped to 21 chromosomes. A stepwise hierarchical sampling strategy with priority based on 4×10^5 SSR data-points was used to construct a core collection from the 23090 initial collections. The core collection consisted of 1160 accessions, including 762 landraces, 348 modern varieties and 50 introduced varieties. The core accounts for 23.1% of the 5029 candidate core accessions and 5% of the 23090 initial collections, but retains 94.9% of alleles from the candidate collections and captures 91.5% of the genetic variation in the initial collections. These data indicate that it is possible to maintain genetic diversity in a core collection while retaining fewer accessions than the accepted standard, i.e., 10% of the initial collections captured more than 70% of their genetic diversity. Estimated genetic representation of the core constructed by preferred sampling （91.5%） is much higher than that by random sampling （79.8%）. Both mean genetic richness and genetic diversity indices of the landraces were higher than those of the modern varieties in the core. Structure and principal coordinate analysis revealed that the landraces and the modern varieties were two relatively independent subpopulaUons. Strong genetic differentiation associated with ecological environments has occurred in the landraces, but was relatively weak in the modern culUvars. In addition, a mini-core collection was constructed, which consisted of 231 accessions with an estimated 70% representation of the genetic variation from the initial collections. The mini-core has been distributed to various research and breeding institutes for detailed phenotyping and breeding of genetic introgression lines.     

Protective effects of nitric oxide on salt stress-induced oxidative damage to wheat (Triticum aestivum L.) leaves

The changes of chlorophyll and malondialdehyde (MDA) contents, plasma membrane permeability confirmed that 0.1 and 1 mmol/L sodium nitroprusside (SNP), a donor of nitric oxide (NO) in vivo, could markedly alleviate the oxidative damage to wheat (Triticum aestivum L.) leaves induced by 150 and 300 mmol/L NaCl treatments, respectively. Further results proved that NO significantly enhanced the activities of Superoxide dismutase (SOD) and catalase (CAT), both of which separately contributed to the delay of O ₂ ⁻ and H₂O₂ accumulation in wheat leaves under salt stress. Meanwhile, the accumulation of proline was apparently accelerated. Therefore, these results suggested that NO could strongly protect wheat leaves from oxidative damage caused by salt stress.     

Molecular identification of asymmetric somatic hybrids between wheat and Italian ryegrass

Sexual incompatibility between common wheat and Italian ryegrass was an obstacle for transferring useful traits from italian ryegrass to wheat. In order to use those desirable genetic resources to improve wheat and to create new cytoplasmic germplasm, the protoplasts of wheat and Italian ryegrass were successfully electrofused and the somatic hybrid plants were regenerated. Examination with 6 restriction enzymes, 13 probes including 9 mtDNA probes (H454, Pst24, B30, Pro I, 490, B342, pHJ2-7-1, B376, 7), 3 cpDNA probes (pHvc p1, pHvc p5 and pHvc p8) and one nuclear DNA probe-- pTA71 (rDNA) in total 73 enzyme/ probe combinations revealed rich polymorphism between the fusion partners. RFLP analysis indicated that approximately 93.4% of the regenerated plants were true somatic hybrids. AFLP analysis implied that the somatic hybrids were highly asymmetric. The RFLP analysis using mt- and cpDNA specific probes also demonstrated the non-coexistence of mitochondria and chloroplasts from the fusion partners in the somatic hybrid cells.     

小麦与高冰草体细胞杂种F5代麦谷蛋白及SDS沉降值分析

通过对普通小麦与高冰草体细胞杂种F5代 175个株系的麦谷蛋白及SDS沉降值分析 ,发现杂种F5代有 10种不同的麦谷蛋白组成 ,并出现了 1Ax1,1Ax2 ,1Bx13,1By16 ,1By8,1Dx5等 6种不存在于亲本小麦中的高分子量麦谷蛋白亚基及 1Bx13 1By16 ,1Bx7 1By8,1Dx5 1Dy12等 3种新亚基组合 ,部分杂种中出现了高冰草的麦谷蛋白特征谱带 ;杂种株系的SDS沉降值表现出较大的差异 ,其中一些杂种的SDS沉降值明显高于亲本普通小麦 .结果表明 ,通过体细胞杂交有可能将野生禾草的优良性状引入普通小麦 ,创造出多种不同组成及组合的新种质 ,为小麦品质育种开辟新途径     

干旱和CO2浓度升高对不同春小麦光合作用和气孔阻力及水分蒸腾效率的影响

为考察春小麦不同抗旱性品种对干旱和高CO2反应的差异,在不同土壤持水量和CO2浓度梯度条件下,对高产性高原602和抗旱性定西24两种春小麦交叉处理一个生长季,结果发现CO2浓度倍增,使两种春小麦的光合速率、气孔阻力和水分蒸腾效率显著增加,增加幅度与土壤含水量分别为正、负和正相关.这种变化幅度在抗旱性不同的春小麦之间进行比较,尤其是在干旱条件下,发现春小麦光合作用、气孔阻力和水分蒸腾效率的变化幅度,抗旱性强的春小麦表现出更有利于生长的趋势.这说明春小麦对CO2浓度升高的反应存在着品种间的差异,也就是说不仅和土壤持水量相关而且还与春小麦抗旱性有关.     

长期继代小麦培养细胞的染色体结构变异特征

本文对继代培养12．5年的普通小麦济南177愈伤组织染色体的结构变异进行了研究，并与继代时间为1．5－8．6年的进行比较，发现其染色体结构变异的类型发生了改变，除了原有的少数染色体断片和双着丝粒染色体以外，较高频率地出现了近端着丝粒染色体和端着丝粒染色体，分布频率分别为91．11％和84．44％。同工酶和RAPD分析表明，不同继代时期的培养细胞基因的组成和表达存在差异。     

两种小麦的原生质体培养再生植株

报道两种基因型小麦昌乐5号和山农587胚性悬浮细胞系的建立和原生质体再生植株。小麦昌乐5号和山农587的胚性愈伤组织继代一年以后形成部分颗粒状和粉粒状结构,可用于悬浮培养。当分散好、生长快的胚性悬浮细胞系形成后,即可作为原生质体培养的材料。较长时间的悬浮培养容易使材料的胚性丧失,昌乐5号的胚性悬浮细胞系20天左右建成,其原生质体再生植株的频率为14/10~5(再生植株数/植板的原生质体数);而山农587的悬浮培养物需5个月以后才能用于原生质体培养,其原生质体再生植株的频率仅为4/10~6。     

八倍体小偃麦远中2染色体组构成的研究

利用来源于四倍长穗偃麦草的六倍体小偃麦８８０１与八倍体与小偃麦远中２杂交，通过Ｆ１芬粉母细胞减数分裂染以体行为分析，发现杂种Ｆ１中染色体组主要构型为１４Ⅱ＋２１Ⅰ，平均构型为：０．８９Ⅲ＋１２．５３Ⅱ＋２１．０７Ⅰ，从而确认亲本八倍体小偃麦远中２染然体组中不含有偃麦草的Ｅ组染色体。     

激素对小麦幼穗组织培养效果的影响研究

分析了激素对两个基因型小麦幼穗组织培养效果的影响,结果表明,愈伤组织诱导率在每个组合中均可达100％,诱导培养基中加入2,4-D对植株再生和绿点分化频率的效果要显著优于Picloram,在所采用的有效2,4D浓度范围内,随着浓度的升高,绿苗的再生频率呈现上升的趋势,2,4-D的量为4.5mg／L时,植株的再生频率平均可以达到45％左右。