【作者】 赵万春；

【导师】 张改生；

【作者基本信息】 西北农林科技大学 ， 作物遗传育种学， 2010， 博士

【摘要】 簇毛麦为异株授粉一年生或多年生二倍体牧草,主要分布于地中海东北部地区,它抗小麦多种主要病害,而且具有耐寒,分蘖力强,生长繁茂,多小花,籽粒蛋白质含量高,耐盐抗旱等特性。位于簇毛麦的1V染色体上的高分子量谷蛋白基因(Glu-V1)、贫硫(ω-型)和富硫(γ-型)醇溶蛋白基因(Gli-V1)和低分子量醇溶蛋白基因(Glu-V3)是改良小麦品质的宝贵基因资源。本研究利用分子标记和细胞学技术创制、筛选和鉴定了2个新的小麦-簇毛麦整臂互补易位系T1DS?1VL和T1DL?1VS,并评估了其对小麦抗病性和籽粒品质的影响,旨在为小麦改良提供新的种质资源。结果如下:1、在小麦第1部分同源群染色体的短臂(1DS)和长臂(1BL)的端部和近着丝粒区域选择设计了96对EST-STS引物,成功开发出了可以检测小麦背景中簇毛麦1V染色体臂的4个EST-STS分子标记。位于1DS上的2个标记BE499250-STS和BE591682-STS/RSAⅠ为共显性标记,可以扩增出1条1VS的特异片段,同时可以扩增出1条1DS特异片段,能将1DS和1AS、1BS区分开来,因此用这2个标记不仅可以检测小麦背景中簇毛麦1V染色体的短臂,还可以检测小麦1D染色体的短臂。而位于1BL上的2个标记BE518358-STS/HAEⅢ和BE585781-STS/RSAⅠ是显性标记,分别扩增出2条1VL和1条1VL的特异片段,但扩增的小麦染色体片段不能区分1DL与1A和1B。2、以中国春的1D单体为母本与中国春-簇毛麦1V染色体的二体附加系(DA1V)杂交。F1植株的根尖制片,计数并选择有42条染色体的植株(为1D和1V的双单体)自交形成F2群体,用我们开发的可追踪1VS和1VL的EST-STS标记筛选282株F2个体,并经基因组原位杂交(Genomic in situ hybridization,GISH)鉴定出了5个杂合互补的Robertsonian易位株和1个非Robertsonian易位株。通过将易位染色体的C带和GISH形态与根尖细胞有色分裂中期1D和1V染色体的C带和GISH形态比较,确认在5个杂合互补的Robertsonian易位株中,3株为杂合互补的T1DS?1VL易位系(08-46-7、08-46-123和08-46-208),2株为杂合互补的T1DL?1VS易位系(08-46-56和08-46-83)。从52株来源于杂合易位株T1DL?1VS83的F3植株中鉴定获得2株纯合互补的T1DL?1VS易位系;从68株来源于杂合易位株T1DS·1VL208的F3植株获得4株纯合互补的T1DS?1VL易位系。3、中国春-簇毛麦的T1DS?1VL和T1DL?1VS易位系抗病性鉴定结果表明,2个新易位系对白粉病的抗性水平和中国春的抗白粉病水平相同,都属于中抗。但2个易位系对4个条锈病小种(条中31号、条中32号、条中33号和水源11致病类型4)的抗性与中国春的不同。易位系T1DL?1VS对4个条锈病小种均为中感或高感,发病程度较中国春严重;而易位系T1DS?1VL对条中31号中抗、对条中32号中感、对条中33号和水源11-4免疫,发病程度较中国春轻。因此,在簇毛麦1VL上可能含有抗条锈病基因。4、中国春-簇毛麦T1DS?1VL和T1DL?1VS易位系的籽粒蛋白质含量与中国春小麦的籽粒蛋白质含量差异并不显著;但是易位系T1DS?1VL的Zeleny沉淀值(10.0 ml)极显著低于中国春的Zeleny沉淀值(30.7 ml),形成时间和稳定时间为1.5 min和1.2 min,分别比中国春形成时间(4.2 min)和稳定时间(4.2 min)减少了2.7 min和3.0 min,其面团弱化度和粉质质量指数为199 FU和19,分别比中国春的弱化度(98 FU)和粉质质量指数(64)增加了101 FU和减少了45;而易位系T1DL?1VS的Zeleny沉淀值(37.4 ml)极显著高于中国春,形成时间为5.0 min,稳定时间为5.9 min,比中国春的形成时间和稳定时间分别增加了0.8 min和1.7 min,其面团弱化度和粉质质量指数分别为47 FU和86,分别比中国春的弱化度和粉质质量指数降低了51 FU和增加了22。结果说明T1DS?1VL易位降低小麦面筋强度和品质;而T1DL?1VS易位增强面筋强度,改善小麦品质。推测簇毛麦的高分子量谷蛋白亚基基因Glu-V1可能位于1VS。更多还原

【Abstract】 Dasypyrum villosa,also named Haynaldia villosa,is an allogamous annual or perennial diploid grass. It is native to the north-eastern part of the Mediterranean region. Haynaldia villosa possesses many important agronomic traits,including resistance to many main wheat diseases,as well as winter hardiness,vigorous tilling ability,multi-spikelets,high grain protein content,salt and drought tolerance. On chromosome 1V there are genes at complex loci coding for high molecular weight glutenins (Glu-V1),sulfur-poor (ω-type) and sulfur-rich (γ-type) monomeric prolamins (Gli-V1),and low molecular weight (LMW) polymeric prolamin proteins (Glu-V3). All these genes are very precious genetic resources for improving wheat quality. In the present study,we used molecular markers and cytogenetic techniques to develop and identify two new compensating,Robertsonian translocation lines T1DL?1VS and 1DS?T1VL,and evaluate the effects of this translocation on wheat diseases and grain quality. The results are as following:1、From 48 pair bin-mapped EST (expressed sequence tags) primers on chromosome 1DS and chromosome 1BL respectively were selected from the wheat EST,24 markers from the centromeric bin and 24 markers from the telomeric bin. Four EST- STS (sequence target site) markers were developed. Two markers BE499250-STS and BE591682-STS/RSAⅠlocated on short arm are co-dominant markers,can trace a polymorphic segment on 1VS,and a 1DS fragment amplified by this marker could be distinguished from 1A and 1B. The other two markers BE518358-STS/HAEⅢand BE585781-STS/RSAⅠlocated on long arm produced dominant markers,the former one can amplified two 1VL fragments and later one can detect one 1VL segment,where the 1DL band co-migrated with those of 1A and 1B.2、Using the monosomoic 1D line as female parent crossed with the disomic chromosome 1V addition line DA1V. Make slides with the root taps of F1 plants,count and select the plants with 42 chromosomes which must be double monosomic plants ( 2n=42,20Ⅱ+Ⅰ1 D+Ⅰ1V) ,and keep these plants self-cross and produce F2 population. A total of 282 F2 plants derived from F1 plant with 2n=42 chromosomes were screened with the four STS markers. 36 plants only showed special polymorphic bands for the two short arm markers,while 27 plants only showed special polymorphic bands for the two long arm markers. 5 heterozygous Robertsonian translocation plants and 1 heterozygous non-Robertsonian translocation plants were identified by GISH. According to the C-banding and GISH patterns of 1D and 1V chromosomes and the critical chromosomes involved in wheat-H.vallosa compensating Robertsonian translocation lines at mitotic metaphase,Of the 5 Robertsonian translocations , 3 T1DS?1VL heterozygous compensating Robertsonian translocation plants(08-46-7、08-46-123 and 08-46-208) and 2 T1DL?1VS heterozygous compensating Robertsonian translocation plant(08-46-56 and 08-46-83) were identified. 2 homozygous T1DL?1VS plants were detected from 52 F3 plants derived from heterozygous translocation plant T1DL?1VS83; while 4 homozygous T1DS?1VL plants were identified by GISH in the 68 F3 plants derived from heterozygous translocation plant T1DS?1VL208.3、Results of resistance to stripe rust and powdery mildew in adult plant for Chinese Spring,T1DL?1VS and T1DS?1VL lines showed,the resistance level to powdery mildew for two new translocation lines T1DS?1VL and T1DL?1VS were similar to CS,all of them were mid-resistance. But resistance to four stripe rust races (CYR31,CYR32,CYR33 and Su11-4) for two translocation lines T1DL?1VS and T1DS?1VL were different with that of CS,plants of the translocation line T1DL?1VS were moderately to highly susceptible to the four stripe rust races,and more serious than CS. Plants of the translocation line T1DS?1VL were moderately susceptible to CYR31 and CYR32,and highly resistant to immune to CYR33 and Su11-4. Therefore,some resistant genes to stripe rust perhaps locate on 1VL.4、The seed protein concentration of CS,two new translocation lines T1DS?1VL and T1DL?1VS were 15.47%,15.20% and 14.81% respectively,and was not significant different. Zeleny sedimentation value (10.0 ml) of the translocation line T1DS?1VL was significantly lower than that of CS (30.7 ml),developing time and stable time were1.5 min and 1.2 min,and decreased 2.7 min and 3.0 min than developing time (4.2 min) and stable time (4.2 min) of CS respectively,weakness of gluten and quality index of farinograph were 199 FU and 19,and increased 101 FU and decreased 45 than that of CS (98 FU and 64) respectively. Zeleny sedimentation value of the translocation line T1DL?1VS was 37.4 ml and was significantly higher than that of CS,developing time and stable time were 5.0 min and 5.9 min,and increased 0.8 min and 1.7 min than that in CS respectively,its weakness of gluten and quality index of farinograph were 47 FU and 86,and decreased 51 FU and increased 22 than that of CS. The results showed that T1DS?1VL translocation decrease gluten strength and do not enhance wheat quality,but T1DL?1VS translocation can increase gluten strength and enhance wheat quality. We speculated that the high molecular weight glutenins (Glu-V1)of Dasypyrum villosa probably located on 1VS.更多还原