SSCP Marker Development and Analysis of Candidate Restorer Gene Rf1 for Cotton Cytoplasmic Male Sterility

[Objective] Locating the cotton cytoplasmic male sterility (CMS) restorer gene Rf₁ is important for investigating restorer gene mechanisms and improving restorer lines. In our previous study, a gene cluster, with nine Pentatricopeptide repeat(PPR) genes and nine other genes, was found within the 160-kb Rf₁ target region in Scaffold 333. The objective here was to improve the density of Rf1-linked markers in the target region and determine the expression profiles of candidate genes. [Method] Using the sequences of the 18 genes, we designed 155 single-strand conformation polymorphism (SSCP) primers covering all of the gene sequences to identify the polymorphic SSCP markers between the fertile and sterile pools. Additionally, real-time polymerase chain reaction(PCR) was performed to analyze the expression profiles of eight candidate genes in the four developmental stages of buds of sterile, maintainer and restoring lines, respectively. [Result] In total, 15 polymorphic primers were identified. A genotype analysis of the F₂ population was conducted using the 15 primers and 3 other polymorphic simple sequence repeats (SSR) markers. The markers were distributed in a 4.8 cM range. In addition, owing to the influence of sterile cytoplasm or restorer genes, most of the genes showed different expression patterns in the four developmental stages of the three lines' buds. [Conclusion] SSCP markers tightly linked to Rf₁ were identified and the expression profiles of candidate genes were determined. This study provides a basis for the further fine mapping of restorer genes and for candidate gene screening.     

Genomewide association analysis of salt tolerance in soybean [Glycine max (L.) Merr.]

Salinity is a common abiotic stress causing soybean [Glycine max (L.) Merr.] yield loss worldwide. The use of tolerant cultivars is an effective and economic approach to coping with this stress. Towards this, research is needed to identify salt‐tolerant germplasm and better understand the genetic and molecular basis of salt tolerance in soybean. The objectives of this study were to identify salt‐tolerant genotypes, to search for single‐nucleotide polymorphisms (SNPs) and QTLs associated with salt tolerance. A total of 192 diverse soybean lines and cultivars were screened for salt tolerance in the glasshouse based on visual leaf scorch scores after 15–18 days of 120 mM NaCl stress. These genotypes were further genotyped using the SoySNP50K iSelect BeadChip. Genomewide association mapping showed that 62 SNP markers representing six genomic regions on chromosomes (Chr.) 2, 3, 5, 6, 8 and 18, respectively, were significantly associated with salt tolerance (p < 0.001). A total of 52 SNP markers on Chr. 3 are mapped at or near the major salt tolerance QTL previously identified in S‐100 (Lee et al., 2014). Three SNPs on Chr. 18 map near the salt tolerance QTL previously identified in Nannong1138‐2 (Chen, Cui, Fu, Gai, & Yu, 2008). The other significant SNPs represent four putative minor QTLs for salt tolerance, newly identified in this study. The results above lay the foundation for fine mapping, cloning and molecular breeding for soybean salt tolerance.     

抗旱节水小麦分子遗传育种研究进展

干旱缺水等自然灾害会导致小麦产量年度间变幅较大,尤其在小麦主产区黄淮地区更为严重。小麦抗旱节水育种是应对干旱的重大措施。本综述对小麦抗旱节水常规育种、抗旱节水分子遗传育种相关性状QTL定位、抗旱相关功能基因克隆鉴定、转基因等方面的研究进展进行了综述。水旱亲本杂交与异地交叉选择是卓有成效的常规育种方法,通过分子标记鉴定了关于根重、根长、胚芽鞘、高水分利用效率等相关性状的大量QTL;42个抗旱相关基因被克隆并进行基因功能分析和验证,均从不同代谢通路上影响着小麦抗旱性;14个来自不同供体的抗旱相关基因被研究者导入小麦品种后,转基因小麦植株的抗旱能力均得到不同程度的提高,部分植株在产量和其它抗逆性方面也得到提高。以上研究进展为抗旱节水小麦分子设计育种提供了理论依据和发展方向。     

水稻蜡质稀少突变体wax1的鉴定及基因定位

【目的】 蜡质是植物表面的一种重要保护物质,筛选和鉴定水稻蜡质相关突变体有助于解析水稻蜡质形成的遗传机制。【方法】利用EMS诱变籼稻品种湘早籼6号,从突变体库中筛选出一个蜡质稀少的突变体wax1,考查突变体的形态特征和农艺性状,利用突变体与02428杂交的F₂群体定位目标基因,并通过荧光定量PCR分析相关基因的表达情况。【结果】与野生型相比,wax1突变体具有叶片短小皱褶、叶表面蜡质晶体减少、穗长变短等形态特征;在农艺性状方面,突变体的株高、每穗总粒数和千粒重显著降低,但有效穗数显著高于野生型;遗传分析表明,wax1的突变表型受一对隐性核基因控制,将WAX1基因定位在第10染色体上SSR标记RM5806与InDel标记P1之间,物理距离约为49.8 kb;定位区间内的测序结果表明,突变体中β-酮脂酰-CoA合酶的编码基因发生单碱基突变,导致催化活性中心的一个氨基酸发生改变。WAX1基因的突变显著提高了同源异型盒基因OSH6的表达,同时引起部分IAA基因的差异表达。【结论】WAX1基因突变引起叶表面蜡质晶体的减少,同时通过影响茎尖分生组织和生长素的信号转导引起植株生长发育等多方面的异常。     

玉米单倍体高自然加倍材料吉Gjb335DH3的遗传分析与QTL初定位研究

玉米单倍体育种技术加倍率低的问题是单倍体育种技术应用限制条件。本研究以雄穗低自然加倍材料DHL287为母本、雄穗高自然加倍材料吉Gjb335DH³为父本，构建单倍体雄穗高自然加倍群体。根据集群分离分析法（BSA，Bulked segregant analysis），结合SNP标记，使用滑动窗口分析3组BSA混池基因型数据，最终在1、3、4、5、7号染色体上筛选出23个与雄穗高自然加倍相关的QTL位点。其中，3组BSA混池结果都在1、3、4号染色体上定位到4个共有的显著QTL区段，说明4个QTL区段具有很强的重演性。同时，在5、7号染色体上分别定位到3个、2个与单倍体雄穗高自然加倍相关的QTL位点。     

寡核苷酸探针套涂染结合基因组原位杂交和分子标记分析准确鉴定小麦-百萨偃麦草异源易位系的研究

培育小麦异源易位系是转移和利用外源基因的有效途径，快速准确的染色体鉴定方法有利于提高染色体工程效率。本研究以电离辐射诱致的2个普通小麦-百萨偃麦草染色体5J易位系（NAU16YJ127和NAU16YJ124）为例，分析了综合利用本课题组前期开发的寡核苷酸探针套［包括(GAA)₁₀、pSc119.2-1、pAs1-1、pAs1-3、AfA-3和AfA-4六个寡核苷酸探针］涂染，结合基因组原位杂交和分子标记等方法鉴定小麦异源易位系的效果。结果表明，NAU16YJ127（2n=6x=42）包含一对小麦-百萨偃麦草易位染色体T5JS·5JL-5AL#1，同时在染色体6A和2D长臂顶端也发生了明显的相互易位；NAU16YJ124（2n=6x=44）附加了一对小麦-百萨偃麦草易位染色体，同时包含一对6A长臂缺失染色体del6AL，其缺失区段易位到5JL近端部，形成T5JS·5JL-6AL，其他染色体未见明显变化。利用21个5J特异分子标记（19个位于长臂和2个位于短臂）分析发现，两个易位系均含有全部5J短臂标记，同时T5JS·5JL-5AL#1含有12个长臂标记，而T5JS·5JL-6AL仅含有2个长臂标记，确证两个易位系断点不同，并将21个标记定位于4个不同区段，其中位于5J短臂的2个标记位于1个区段，位于5J长臂的19个标记位于3个区段（5JL-1~3），分别包含2、10和7个标记；发现3个为可以同时识别5J和5A的共显性标记，其中NAU16YJ124含有全部5A标记，而NAU16YJ127仅包含其中2个标记，进一步验证了细胞学鉴定结果。本研究结果表明，寡核苷酸探针套涂染、基因组原位杂交和分子标记分析相结合为准确鉴定小麦异源易位系提供了新方法。     

长江流域生物资源及生态环境研究进展

以CNKI平台中的文献数据,利用文献计量学方法,对其文献的增长趋势及期刊分布进行分析,并基于作者和机构合作网络、关键词共现的聚类知识图谱及突变检测等方法,探究我国长江流域生物资源及生态环境主题的研究热点及其研究前沿。研究表明:研究文献总体上呈递增趋势,2016年开始呈现激增态势;作者、机构间的交流合作较少,作者间合作大部分局限在机构内部的研究者之间的合作,合作相对活跃的机构主要有长江科学院、长江流域水资源保护局及长江水利委员会等;其优势学科领域主要为环境科学、区域经济学、水利工程学、农业经济等。当前我国长江流域生物资源及生态环境的研究领域有5个方向:(1)开展长江经济带城市群生态环境评价与保护、城镇化与生态文明建设,以及绿色发展等研究;(2)开展长江三峡与三峡库区的生态环境监测与保护、森林资源养护、水土保持与流失、资源与生态补偿机制研究及其生态环境的可持续发展等研究;(3)开展长江流域自然保护区的生态环境保护与规划、生态保护红线及其生物资源养护与生物多样性保护等研究;(4)在长江大保护背景下,开展长江流域生态环境的保护与修复、流域综合治理,以及水生生物资源的合理开发利用的研究;(5)开展长江源区水电资源开发、水环境保护及其渔业生态环境保护等研究。为科学管理和保护长江流域的重要生物资源,提出建立基于科学设计的全流域生物资源和环境监测系统,建立全流域的生物资源及生态系统的生物数据模型,发展长江流域渔业资源养护与开发的风险评估决策系统,发展长江流域科学的渔业资源增殖放流技术体系,发展基于GIS的长江全流域的渔业资源与环境监测和管理决策信息服务系统等建议。     

Identification and QTL mapping of Z550, a rice backcrossed inbred line with increased grains per panicle

An elite backcrossed inbred line Z550 with increased grains per panicle was identified from advanced backcrosses between Nipponbare and Xihui 18 by simple sequence repeat (SSR) marker-assisted selection (MAS). Z550 carries 13 substitution segments distributed on chromosomes 1, 6, 7, 8, 9, 10, and 12, with an average substitution length of 1.68 Mb. Compared with the Nipponbare parental line, plant height, panicle length, spikelets per panicle, grains per panicle, and grain weight for Z550 were significantly increased. While the grain width of Z550 was significantly narrower, and the seed setting ratio (81.43%) was significantly lower than that of Nipponbare, it is still sufficient for breeding purposes. Quantitative trait loci (QTLs) mapping for important agronomic traits was conducted with the F₂ population derived from Nipponbare crossed with Z550 using the restricted maximum likelihood (REML) method. A total of 16, including 12 previously unreported QTLs were detected, with contribution rates ranging from 1.46 to 10.49%. Grains per panicle was controlled by 8 QTLs, 5 of which increased number of grains whereas 3 decreased it. qGPP-1, with the largest contribution (10.49%), was estimated to increase grains per panicle by 30.67, while qGPP-9, with the minimum contribution rate (2.47%), had an effect of increasing grains per panicle by 15.79. These results will be useful for further development of single segment substitution lines with major QTLs, and for research of their molecular functions via QTL cloning.