【作者】 李曙光；

【导师】 盖钧镒； 赵团结；

【作者基本信息】 南京农业大学 ， 作物遗传育种， 2009， 硕士

【摘要】 杂种优势利用是大幅度提高作物单产的重要途径之一,目前大豆杂种优势利用育种还处于探索阶段,虽然国内多家单位已育成数十个质核互作雄性不育系并实现了“三系”配套,而且利用质核互作雄性不育系,育成3个正式通过品种审定的大豆杂交种品种,但是在生产上尚未大规模推广应用。质核互作雄性不育系及其育性恢复是杂交种制种的基础,为此本研究以N21566和N8855为不育细胞质背景进行新质核互作雄性不育系的选育,以及NJCMS3A雄性育性基因的SSR标记定位,另外对大豆核不育突变体NJS-1H进行遗传分析与细胞学研究。主要结果如下：1.以国家大豆改良中心育成的细胞质雄性不育系早代材料为基础,利用相应的保持系连续回交三代,初步育成以N21566为不育细胞质背景的2个新质核互作雄性不育系NJCMS 4A(BC8F1)与NJCMS (N21566/73-935) (BC3F1);以N8855为不育细胞质背景的3个新质核互作雄性不育系NJCMS5A(BC8F1), NJCMS(N8855/88-48) (BC4F1)与NJCMS (N8855/73-935) (BC3F1)。NJCMS 4A/NJCMS 3A、NJCMS 5A/ NJCMS 1A分别构成同核异质系。这5个质核互作雄性不育系在不同年份间花药散粉性、花粉萌发率和成熟期植株表型等方面鉴定表明不育性表现稳定。2.利用NJCMS3A的细胞质、核供体亲本N21566和N21249配置杂交组合,(N21566/N21249) F1结荚正常可育,对(N21566/N21249)F2代、(N21566/N21249 //N21249)BC1F1代分离群体进行育性鉴定,F2和BC1F1群体的可育与不育株分离比例分别符合3：1和1：1,表明NJCMS3A雄性育性由一对基因控制,其中可育等位基因显性,而与之对应的不育等位基因隐性。选用793对大豆SSR引物对F2和BC1F1群体进行大豆质核互作雄性不育系NJCMS3A雄性育性基因定位,发现O连锁群上的3对SSR引物(Satt331,CSSR133和Satt477)与NJCMS3A雄性育性基因连锁,遗传距离分别为8.1 cM,11.4 cM,13.3 cM。3.通过对核雄性不育突变体NJS-1H育性分离株行中可育单株的衍生后代M8：9,M9：10与Mi10：11的育性鉴定,验证该核不育突变体雄性育性由一对基因控制。进一步细胞学研究发现,该突变体在减数分裂时染色体联会异常,出现单价体。减数分裂过程中出现染色体落后,不对称分裂,不同步分裂等现象；在四分体阶段,出现各种畸形多分体以及大量微核。在花粉粒阶段,花粉粒退化,无内容物。总之,从减数分裂到花粉粒发育至成熟期都有败育发生,但是大量败育主要发生在减数分裂阶段,四分体以后的败育可能是减数分裂染色体联会异常的结果。更多还原

【Abstract】 Utilization of heterosis is one of the most important ways to increase crop yield quickly, however, it is still at primary stage for soybean. Up to now dozens of cytoplasmic-nuclear male-sterile (CMS) lines have been developed and many sets of "three lines" have been obtained in many different research institutions in China, and three soybean hybrid cultivars based on the "three lines" of CMS system have been released. Because of the poor outcrossing rate of the CMS lines under natural cultivation condition, it is very difficult to produce enough soybean hybrid seeds with cheap price for soybean production. The objectives of this thesis were to develop new CMS lines with different sterile cytoplasm sources and nuclear backgrounds, to reveal the genetic mode of CMS line NJCMS3A and map its male fertility gene through SSR marker technique, and to investigate the cytomorphology of nuclear male-sterile mutant NJS-1H in soybean. The main results were as follows:1. Some CMS progenies derived from N8855 and N21566 cytoplasm were obtained from National Center for Soybean Improvement, and five new CMS lines have been developed by three times of successive backcrossing with corresponding maintainer line. Among them, two possessed identical male-sterile cytoplasm (N21566):NJCMS 4A (BC8F1) and NJCMS (N21566/73-935) (BC3F1); the other three ones derived from the male-sterile cytoplasm (N8855):NJCMS5A (BC8F1), NJCMS(N8855/ 88-48) (BC4F1) and NJCMS (N8855/73-935) (BC3F1). On the other hand, NJCMS 4A/NJCMS 3A、NJCMS 5A/ NJCMS 1A were iso-nuclear lines with different sterile cytoplasm. The male sterility of these five CMS lines was stable among different years based on pollen dispersiveness and germination viability, as well as fertility performance at maturity stage.2. The cross between cytoplasmic and nuclear donor parents (N21566/N21249) of CMS line NJCMS3A was made, its F1 plants were fertile according to the pod-setting situation. The results of the fertility performance of (N21566/N21249) F2, (N21566/N21249//N21249) BC1F1 generations showed that the segregation ratio of two phenotypes, i.e. fertile and sterile plants, in the F2 and BC1F1 fit 3:1 and 1:1 expected ratios, respectively. It indicated that there might be one pair of gene for the male fertility of NJCMS3A, and the plants which had the dominant gene were fertile, vice versa, sterile. The above-mentioned F2 and BC1F1 populations were utilized to map male fertility gene of NJCMS3A with 793 SSR markers selected randomly from the soybean public genetic linkage map reported by Song et al.(2004). Three SSR markers, Satt331, CSSR133 and Satt477 in linkage group O, were found linked to the fertility gene Rf of NJCMS3A, the genetic distance were Satt331/8.1 cM, CSSR133/11.4 cM, Satt477/13.3 cM, respectively..3. It confirmed that the male fertility of a nuclear male-sterile mutant NJS-1H was controlled by a single recessive nuclear gene according to the fertility segregation data of the NJS-1H progenies (M8:9, M9:10 and M10:11). Nomal squash and acetocarmine dyeing observation indicated that the meiosis of pollen mother cells of NJS-1H sterile plants was highly irregular with chromosome asynapsis or desynapsis. There were univalent, chromosome lagging, unequal and asynchrony distribution among other phenomena during meiotic division. At tetrad stage, various malformed polyad with micronucleus can be observed, then, the degenerated microspores were empty, shrunken without cytoplasm at uninuclear microspore stage. The results showed that there appeared different sterility characters from initial stages of meiosis to pollen maturity, and the pollen abortion of NJS-1H occurred mainly at meiosis stage, and the disorder of chromosome synapsis at meiosis might cause the pollen degeneration after tetrad stage.更多还原