【作者】 李兴锋；

【导师】 王洪刚；

【作者基本信息】 山东农业大学 ， 作物学， 2003， 博士

【摘要】 以人工合成的八倍体小黑麦劲松49、八倍体小滨麦和八倍体小偃麦中5为材料，在对其染色体构成和性状特点综合评价的基础上创制了小麦-黑麦-滨麦草、小麦-黑麦-中间偃麦草两个三属杂种。对三属杂种及其自交后代的性状特点和细胞遗传学特点进行了分析；并综合利用细胞学、染色体C-分带和染色体原位杂交技术分析了三属杂种后代的染色体分离和传递特点；在两个三属杂种后代进行了种质系的筛选，并对选育的种质材料进行了性状和遗传特点的评价；对育成的种质系山农030-1中白粉病抗性基因性质和来源进行了分析，并利用RAPD技术对其抗性基因进行了分子标记研究。获得了以下主要结果：1．对八倍体小黑麦劲松49、小滨麦、小偃麦中5的性状特点及染色体构成进行了分析，结果表明这三个八倍体中间材料的染色体数目都为2n=56，花粉母细胞减数分裂中期IPMC MI多数细胞形成28个二价体，细胞学稳定性较好。染色体原位杂交和C-分带结果表明劲松49的染色体组成为22对小麦染色体和6对黑麦染色体，6对黑麦染色体为1R、2R、4R、5R、6R、7R；小滨麦的染色体组成为44条小麦染色体和12条滨麦草染色体。 2. 以劲松49为母本，小滨麦和中5分别作父本进行杂交，创制了小麦-黑麦-滨麦草、小麦-黑麦-中间偃麦草两个三属杂种，其中小麦-黑麦-滨麦草三属杂种的创制尚未见研究报道。研究结果证明，利用人工合成的双二倍体材料创制多属杂种是比较有效的途径。对两个三属杂种F1及其自交后代的性状特点调查结果表明，杂种后代变异类型多样，可以从中筛选综合多属特点的种质材料。劲松49/小滨麦自交后代在形态和籽粒性状方面受母本劲松49的影响较大，大多数植株表现倾劲松49的特点；而劲松49/中5自交后代植株分离类型较为丰富，既有偏向于母本劲松49的类型，也有偏向于父本中5的类型，还出现了一些中间类型和新类型。 3．劲松49/小滨麦、劲松49/中5杂种F1花粉母细胞平均染色体构型分别为13.17Ⅰ+20.82Ⅱ+0.37Ⅲ+0.02Ⅳ、14.32Ⅰ+20.28Ⅱ+0.32Ⅲ+0.04Ⅳ，其相对紊乱系数分别为0.34、0.38，显著高于其亲本。在劲松49/小滨麦、劲松49/中5两个杂种F1花粉母细胞后期I都观察到较高频率的落后单价体，四分体中普遍具有微核，两个杂种F1成熟花粉<WP=9>败育率分别为55.9%、59.2%。此外在两个杂种F1小孢子发生和雄配子体发育过程中观察到多种异常现象。 4．以标记的黑麦和滨麦草DNA为探针，对劲松49/小滨麦杂种F1花粉母细胞和小孢子进行原位杂交分析，发现劲松49/小滨麦杂种F1含有以单价体形式存在的6条黑麦染色体和6条滨麦草染色体，表明亲本劲松49和小滨麦的外源染色体都已传递给杂种F1。在小孢子时期的原位杂交结果表明，黑麦的染色体主要存在于小孢子细胞核中，而滨麦草的染色体多以微核的形式散布在小孢子内。5．劲松49/小滨麦杂交后代中，二价体频率相对较高，多价体频率相对较低；劲松49/中5杂交后代的染色体构型比较复杂，二价体频率相对较低，而多价体频率较高。两个三属杂种后代中，随着自交世代的增进，染色体数目逐渐减少，劲松49/中5自交后代染色体数目降低较快。6．利用染色体C-分带和原位杂交技术对两个三属杂种自交后代进行分析，结果表明在劲松49/小滨麦自交后代中除小麦的染色体外，仅检测到黑麦染色体，大多数植株含有8-12条黑麦染色体，在所观察的后代植株中没检测到滨麦草染色体。在劲松49/中5自交后代的检测中发现，除了小麦、黑麦染色体以外，中间偃麦草的染色体在后代中也得到了保留。综合三属杂种后代的性状特点和染色体分离、传递特点结果，推测偃麦草、黑麦和滨麦草染色体组与小麦染色体组的同源性为：偃麦草＞黑麦＞滨麦草。7．在劲松49/小滨麦后代筛选出种质系山农030-1，根尖细胞有丝分裂及PMC MI染色体构型分析表明其染色体数目为2n=42，减数分裂中期I形成21个二价体，遗传较为稳定。白粉病鉴定结果表明山农030-1对白粉病免疫。原位杂交结果发现山农030-1不含有来自滨麦草的遗传物质，是一个由黑麦的1R染色体短臂与小麦染色体易位形成的小麦—黑麦易位系。经进一步利用染色体C-分带方法对其分析，证明山农030-1为1RS·1BL易位系。在劲松49/小滨麦F6代筛选到种质系山农L050，根尖细胞染色体计数结果表明其染色体数目为2n=56，PMC MI染色体平均构型为1Ⅰ+27.3Ⅱ+0.12Ⅳ，细胞学基本稳定，并且对白粉病免疫。染色体原位杂交和染色体C-分带结果表明山农L050含有5对黑麦染色体，分别为1R、2R、5R、6R、7R。<WP=10>在劲松49/中5 F5代选育出三个染色体数目为42，PMC MI多形成21个二价体的株系，分别命名为山农L077、山农L097、山农L0100。这三个株系的农艺性状较好，遗传也较稳定。8．为确定山农030-1的白粉病抗性基因性质，分别以感病普通小麦中国春、辉县红与山农030-1杂交，配制两个杂交组合山农030-1/中国春、山农030-1/辉县红。根据杂种F1及F2代分离群体的抗病性调查结果，证明山农030-1所含白粉病抗性可能由显性单基因控制，结合F2代原位杂交结果，推断其白粉病抗性可能来自于黑麦的1R染色体。选用200个十聚体核苷酸随机引物对山农030-1/辉县红F2分离群体进行RAP更多还原

【Abstract】 Two trigeneric hybrids were synthesized by crossing amphiploid Triticale(Jinsong49) with Tritileymus and Trititrigia(Zhong5), the morphology and cytogenetic characters of the trigeneiric hybrids and their derivatives were characterized by C-banding and genome in situ hybridizations.. In the selfed progenies of two trigeneric hybrids, 5 germplasms with good traits were obtained. The character and origination of the resistant gene to powdery mildew in Shannong030-1 were analysised, and the RAPD markers of this resistant gene were constructed. Results were as follows：1. The morphological trait and cytogenetic characters of octoploid Jinsong49,Tritileymus, Zhong5 were identified, they all had the chromosome number of 2n=56, and could form 28 bivalent at PMC MI. Chromosome constitution of Jinsong49 were confirmed by C-banding and in situ hybridization, it had 22 pair of wheat chromosomes and 6 pair of rye chromosomes as 1R,2R,4R,5R,6R,7R. Results of in situ hybridization indicated that Tritileymus had 44 wheat chromosomes and 12 Leymus mollis chromosomes.2. Use the Jinsong49 as female parent, pollinated with Tritileymus and Zhong5, two trigeneric hybrids as Triticum-Secale-Leymus, Triticum-Secale-Elytirgia were obtained. The morphological characters of the trigeneric hybrids and their progenies were studied, it showed that the selfed derivatives of Jinsong49/Tritileymus were inclined to Jinsong49. While the progenies of Jinsong49/Zhong5 showed extensive segregation in morphology, some new type and mediate phenotype were observed.3. The mean of chromosome configuration in Jinsong49/Tritileymus F1, and Jinsong49/ Zhong5 F1 were 13.17Ⅰ+20.82Ⅱ+0.37Ⅲ+0.02Ⅳ and 14.32Ⅰ+20.28Ⅱ+0.32Ⅲ+0.04Ⅳ respectively，and their relative chaotic coefficient were 0.34 and 0.38,they were higher apparently than their parent.In the course of microsporagenesis and male gametophyte development, high frequency of laggard can be observed at meiotic AI, as well as the micronucleoli in tetrad. The sterile rate of mature pollen in two trigeneric hybrids were 55.9% and 59.2% respectively. And some abnormal phenomenon were observed4. Using labelled genome DNA of Secale cereale and Leymus mollis as probes，in situ hybridizations showed that there were 6 Secale and 6 Leymus chromosome in the Jinsong49/TritileymusF1, and the GISH pattern of microspores showed that some of rye chromatins exist in the nucleus, some exist in the cytoplasm as micronucleoli, while most Leymus chromatin present in the microspore as the micronucleoli.5. In the selfed derivatives of two trigeneric hybrids, with generation increased, the mean<WP=12>of chromosome number tends to decreased, the chromosome number of progenies from Jinsong49/Zhong5 seems to decrease faster. The results also indicated that the homologous pairing way，and the speeds of chromosome number decreased in the progenies of these two trigeneric hybrids were different.6. Using labelled genome DNA of Secale cereale and Leymus mollis as probes ，genome DNA of Chinese spring as blocked DNA, in situ hybridization were applied to analysis the chromosome constitution of the progenies from Jinsong49/Tritileymu. The results showed most selfed derivative plant contained 8-12 rye chromosomes,but Leymus chromosome was not observed. Analysis of chromosome constitution on germplasm lines L049-5 and L050-1 by using C-banding techniques showed that 5 pair of rye chromosome of them were 1R,2R,5R,6R, 7R.The chromosome constitutions of the progenies from Jinsong49/Zhong5 were identified by C-banding and in situ hybridization, and it was found that chromosomes of rye and Elytrigia existed simultaneously in the selfed derivative plant. And germplasm line W1044-2 had one pair of 1R chromosome and one pair of Elytrigia chromosome, germplasm line W1037-1 had one pair of Elytrigia chromosome and had no rye chromosomes.7.In the progenies of Jinsong49/Tritileymus, a stable germplam line named Shannong 030-1 were obtained. Its chromosome number was 2n=42, and could form 21 bivalents at PMC MI. Resistance ide更多还原