【作者】 刘海英；

【导师】 陈彦惠；

【作者基本信息】 河南农业大学 ， 作物遗传育种， 2012， 硕士

【摘要】 种子活力在农业生产中发挥着的重要作用。农业生产的发展对种子质量的要求越来越严格，要求种子要有更高的播种品质。在种子耐藏性方面，选择高活力种子可以较好地抵抗各种贮藏逆境，有利于种质资源的长期保存。本研究以豫82与沈137为亲本构建的201个重组自交系（RIL）为材料，在人工气候室中进行标准发芽试验、低温发芽试验和人工加速老化发芽试验。用SSR分子标记构建了遗传连锁图谱，采用复合区间作图法（CIM）对玉米种子的发芽势、发芽率、发芽指数、活力指数、苗长、幼苗干重以及根干重等活力相关性状进行了QTL定位以及效应分析。同时，对不同温度环境下和不同人工老化处理条件下种子活力相关性状的QTL及其与环境互作进行了上位性效应分析，以期找到控制种子活力相关性状的稳定的主效基因或QTL,以便利用生物信息学的方法和定位结果对控制种子活力相关性状的候选基因进行预测，开发出与目标性状紧密连锁的分子标记，从而为玉米种子活力相关性状的基因克隆、分子标记辅助选择育种提供重要的参考依据。主要实验和研究结果如下：1．选取能够覆盖玉米全基因组的654对SSR引物在亲本间进行多态性筛选，从而获得了175对多态性好、带型清晰的引物。构建了一张含有171个标记的玉米连锁图谱，覆盖玉米10条染色体，第1、2、3、4、5、6、7、8、9、10染色体长度分别是320.4、263.9、287.5、201.7、288、224.4、122.9、160.4、265.4和216cM，标记数目分别是23、17、17、19、22、16、10、14、17和16。图谱总长度2350.6cM，标记平均间距13.7cM。2．在适温28℃和低温18℃两种温度环境下分别进行标准发芽试验和低温发芽试验，共定位到与种子活力相关的10个QTL，分布于第1、3、4、9和10等5条染色体上，各QTL的贡献率介于5.6%～15.6%之间，其中第3染色体上控制活力指数的qVI3具有最大贡献率为15.6%。在标准发芽试验（28℃）下，控制发芽指数和活力指数的qGI1和qVI1分别位于标记区间phi039-umc2029和umc2029-bnlg1268内。在低温发芽试验（18℃）下，检测到同时控制发芽指数和活力指数一个共同的QTL，即qGI10和qVI10，位于标记区间umc1677-umc1993内，在第10染色体上的159.1cM位置上。相对于其它性状指标，发芽指数和活力指数可以作为种子活力的有效指标。3．在人工加速老化处理2天、4天和6天条件下，共检测到与种子活力相关性状的QTL30个，分布在第1、2、3、4、5、8、9和10染色体上，贡献率在6.3%～12.6%之间。在第10染色体上的157.1cM位置上，检测到一个控制活力指数的稳定QTL，即qVI10，位于标记区间umc1648-umc1667内，可以作为一个分子标记应用于分子标记辅助选择。在第10染色体上发现一个控制种子活力的重要染色体区段，位于10.04-10.06位点，标记区间umc1272-umc1993内。在老化处理2天条件下，控制发芽势、发芽指数和活力指数三个QTL，即qGET2、qGIT2和qVIT2均位于第2染色体上，位于相同的标记区间umc1923-umc1637内。在老化处理4天条件下，在第10染色体上，分别检测到1个控制发芽率和1个控制活力指数的QTL，即qGPF10和qVIF10，位于相邻的标记区间内。在老化处理6天下，控制发芽势、发芽率、发芽指数和活力指数的QTL均在第10染色体上被检测到，qGPS10、qGIS10和qVIS10都位于标记区间umc1648-umc1667，位置相同，可能是一个QTL，表现出一因多效的作用。4．在不同温度环境下，检测到了4对影响种子活力性状的AA上位性效应，且均达到了极显著水平。这4对QTL均表现为非加性效应，但都共同表现为加加上位性效应，表明上位性作用在不同温度环境下对种子活力起着重要作用。在不同老化处理条件下，共检测到11个与种子活力相关的QTL加性效应及加性×环境（AE）互作效应，2对与种子活力相关的AA上位性效应，分别位于第1、3、4、8和10条染色体上，贡献率在2.49%-7.21%之间，控制发芽势的qGE1-3贡献率最大。在第1染色体上的qGE1-3、qGR1-3、qGI1-3、qVI1-3均在标记区间umc1568-bnlg1007内，位置相同或相近，可能是一个基因或表现为紧密连锁，增效基因均来自豫82。在第8染色体上umc1121-mmc0181标记区间内，同时检测到控制发芽势、发芽率、发芽指数和活力指数的QTL，即qGE8-9、qGR8-9、qGI8-9和qVI8-9，增效基因均来自沈137。5．在不同天数老化处理下，亲本豫82和亲本沈137在不同老化处理后，表现出不同的抗老化性，亲本豫82在发芽势、发芽率、发芽指数、活力指数和苗长等种子活力相关性状上均比亲本沈137表现出较强的抗老化性。利用RILs群体定位的结果表明，在老化处理初期，两亲本对种子活力相关性状都有不同的增效作用。但是，随着老化时间的延长，增效基因越来越多的来自活力较强的亲本豫82。因此，在配制杂交组合时，利用豫82做亲本，再通过分子标记辅助选择，利用杂交、回交等育种手段，就可以配制出强优势的杂交组合。更多还原

【Abstract】 Seed vigor plays an important role in the agriculture production. Thehigher sowing seeds quality is required with the development of the agricultureproduction. In the seeds storage, the higher seed vigor is better able to resist all kinds ofadversity, so it’s helpful for long-term preservation of germplasm resource. In this study,201recombinants inbred lines were constructed by Yu82and Shen137, standardgermination test, low temperature germination test and artificial accelerated aginggermination test were carried out in the artificial climate chamber. A genetic map wasconstructed by SSR markers. QTL analysis for germination energy, germinationpercentage, germination index, vigor index, seedling length, seedling dry weight and rootdry weight related traits of seed vigor were conducted by composite interval mapping(CIM) method. The interaction among detected QTLs and the interaction with QTL andenvironment were analysis in different temperature environments and different agingtreatments in order to find stable effect genes or QTL, and candidate genes wereforecasted using the method of bioinformatics. The results will help for gene cloning andmarker assisted section of seed vigor. The main results were as follows:1. In total,654SSR primers were choose to screen polymorphism between Yu82andShen137,175great differences in polymorphism makers were found, and a maize geneticlinkage map contains171markers was constructed which covered10chromosomes ofmaize. The each length of chromosome is320.4,263.9,287.5,201.7,288,224.4,122.9,160.4,265.4and216cM, and the markers of each chromosome are23,17,17,19,22,16,10,14,17and16, respectively. The total genetic distance is2350.6cM with13.7cMaverage distance between markers.2.10QTLs of seed vigor were detected under the standard germination test (28℃)and low temperature germination test (18℃), which distributed on1,3,4,9and10chromosomes, the contribution of each QTL between5.6%and15.6%. The qVI3whichcontrol vigor index has a maximum contribution rate of15.6%. In standard germination,qGI1and qVI1which control germinaton index and vigor index was located in markphi039-umc2029interval and umc2029-bnlg1268interval, respectively. In the lowtemperature germination test, a common QTL which control germination index and vigorindex was detected on chromosome10in159.1cM, which in mark umc1677-umc1993interval. The germination index and vigor index can be used effective traits of seed vigorcompared to other traits of seed vigor.3.30QTLs of seed vigor were detected in the artificial accelerated aging treatments of2days,4days and6days, which distributed on1,2,3,4,5,8,9and10chromosomes,and the contribution rate between6.3%and12.6%. Stability QTL controled vigor indexwas detected in157.1cM on chromosome10, which located in markerumc1648-umc1677interval inside, can be used as a molecular marker in the molecularassistant selection. On the chromosome10, we found an important chromosome segmentsthat controled seed vigor, which located in10.04-10.06points, in umc1272-umc1993mark interval inside.After2days aging treatments, qGET2, qGIT2and qVIT2controled germinationerengy, germination index and vigor index, respectively, were located on chromosome2,in the same mark interval umc1923-umc1637inside. After4days aging treatments,qGPF10and qVIF10controled germination percentage and vigor index, respectively,were located on chromosome10, in neighboring mark interval. After6days agingtreatments, the QTLs controled germination energy, germination percentage, germinationindex and vigor index were located on chromosome10, in the same mark intervalumc1648-umc1667, may be a QTL, showed pleiotropy or tight linkage.4. Four pairs AA which influced related traits of seed vigor were detected indifferent temperature environments, they have reached a significant level. Theyperferenced a non-additive effect, but they were common for AA epistatic effect. Soepistatic effect palys an important role in seed vigor under the different temperatureenvironments. In different aging treatment conditions,11additive effect and additive×environment (AE) interaction effect QTLs were detected, and2pairs AA epistatic weredetected, which distributed on1,3,4,8and10chromosome, the contribution ratebetween2.49%~7.21%, and qGE1-3controled germination energy showed the biggestcontribution rate.On chromosome1, qGE1-3, qGR1-3, qGI1-3, qVI1-3all in the markumc1568-bnlg1007interval, within the same or similar position, it might be a gene orperformance as tight linkage, and the efficiency gene from Yu82. In umc1121-mmc0181mark interval on chromosome8, qGE8-9, qGR8-9, qGI8-9and qVI8-9controledgermination energy, germination percentage, germination index and vigor index weredetected simultaneously, and the efficiency gene from Shen137.5. In different days under the aging treatment, the parents of Yu82and Shen137perferenced different anti-aging, Yu82were better than Shen137in the aspects ofanti-aging, such as germination energy, germination percentage, germination index, andvigor index and seedling length of seed vigor. The results of QTL detected using RILsshowed that two parents had both a synergy to the related traits of seed vigor in the earlyaging treatment; however, more and more efficiency gene came from Yu82with theincrease aging time. Therefore, in the process of mading cross combination, using Yu82as parent inbred line, through some breeding method such as molercular assistant selection, cross and backcross, we can make a strong advantage of hybrid combination.更多还原