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.     

Dynamics of Cotton Rhizosphere Bacterial Community Structure in Cotton Continuous Cropping Field Soil

[Objective] The aim of this study is to understand the dynamics of cotton rhizosphere bacterial community structure in cotton continuous cropping field soil. [Method] 16S rDNA genes were sequenced by high-throughput sequencing technology to determine the community structure of cotton rhizosphere bacteria in different developmental stages using an upland cotton cultivar (Gossypium hirsutum cv. TM-1). [Result] Four dominant phyla were found in the cotton rhizosphere bacterial community including Proteobacteria, Acidobacteria, Planctomycetes, and Bacteroidetes. The four dominant phyla and Firmicutes were largely influenced by cotton root. The relative abundances of Acidobacteria, Planctomycetes, Bacteroidetes were promoted, and Proteobacteria and Firmicutes were inhibited, by cotton root. There were significant differences in community structure, but not species richness or α-diversity among different developmental stages of the cotton rhizosphere bacterial community; the differences between the flowering stage and the budding stage were greater than the differences between the budding stage and the seedling stage. The diversity of the rhizosphere bacterial community in cotton continuous cropping field soil was significantly higher than that of the bulk soil; the β-diversity values of both the rhizosphere and bulk soil bacterial communities were highest in the flowering stage. [Conclusion] The structure and dynamics of the cotton rhizosphere bacterial community in cotton continuous cropping field soil was defined by high-throughput sequencing. The effect of cotton on the rhizosphere bacterial community structure was most significant in the flowering stage.     

Potassium accumulation,partitioning, and remobilization in high-yield spring maize in Northeast China

Potassium (K) plays an important role in maize yield, but K accumulation characteristics in high yielding maize is still not well documented. A two-year field experiment was conducted to investigate the K accumulation characteristics of high yield (HY)（>15 t ha^?1）spring maize compared with medium yield (MY)（10–15 t ha^?1), and low yield (LY)（<10 t ha^?1). The maximum K accumulation stage in maize appeared between V6 and V12 stage (LY, MY, and HY was 125.46, 138.05, and 146.22?kg ha^?1, respectively). Most of the K (94.27–97.13%) was accumulated during vegetative stages. HY exhibited a significantly higher K accumulation than either MY or LY. For different organs, the K remobilization amount of leaf was the highest, and the remobilization amount of stalk was the lowest, the K remobilization amount of leaf and stalk showed as HY?<?MY?<?LY, same as the total K remobilization amount, but the K remobilization amount of sheath and husk plus cob showed the opposite order. These results indicated that sufficient nutrient supply for maize can not only accumulate more K but also delay leaf senescence and maintain high photosynthetic activity, resulting in reduced K remobilization from vegetative organs, and reduced K loss in whole plant.     

Straw mulching with fertilizer nitrogen: An approach for improving crop yield,soil nutrients and enzyme activities

Field experiments were conducted to study soil properties, soil enzymes activities, water use efficiency (WUE) and crop productivity after six years of soya bean straw mulching in the semi‐arid conditions of China. The experiment included four treatments: CK (Control), N (240 kg N ha^‐1), H (soya bean straw mulching at half rate 700 kg ha^‐1 with 240 kg N ha^‐1) and F (soya bean straw mulching at full rate 1,400 kg ha^‐1 with 240 kg N ha^‐1). Soil organic carbon (SOC), soil labile organic carbon (LOC), soil available N (AN), available P (AP) and enzyme activities were analysed after wheat harvesting in 2016 and 2017. Results show that straw amounts had positive effects on the soil fertility indices being higher for treatment F. The SOC, LOC, AN, AP and enzyme activities (i.e. saccharase, urease and alkaline phosphatase) were in the order of F > H > N > CK. High wheat grain yield and WUE were observed for F treatment. A total of six years mulching along with 240 kg ha^‐1 nitrogen fertilizer application is sufficient for wheat yield stability and improving soil properties except urease activities in the semi‐arid condition of China. However, the straw mulching amount should be further studied with minimum nitrogen fertilizer for an environment‐friendly and effective approach for improving the soil biological properties with adequate crop production on a sustainable basis in the semi‐arid region of China.     

“三系”杂交棉组合F1花粉育性与产量和品质的相关性研究

细胞质雄性不育“三系”（不育系、保持系和恢复系）制种因简化制种和节约劳动力成本已成为棉花杂种优势利用的重要手段。研究棉花花粉育性及“三系”组合F1可育花粉率与产量、品质性状的相关性，可以达到强化“三系”制种，提高育种效率的目的。用联苯胺-甲萘酚法测定12个不育系与4个恢复系杂交所得的48个F1的花粉育性，统计了可育花粉率并分析了可育花粉率与产量、品质的相关性。结果表明：－20℃保存条件下，花粉育性随贮藏时间的增加而减弱，51 h内其花粉育性依然保持88%以上，51 h内花粉育性无显著性影响。F1的可育花粉率均低于其恢复系及常规杂交种，大部分组合降低水平在20百分点以内。不同恢复系的恢复力有明显差异, 其中H46恢复力较强。F1的可育花粉率与单株铃数呈显著正相关。本研究结果有助于了解离体花粉的育性变化规律及“三系”杂交组合后代可育花粉率与产量品质的关系，可应用于异地授粉并在一定程度上指导“三系”杂交种选育，为育种工作提供一定的参考。     

上海半自然森林更新苗物种组成与多样性研究

城市半自然森林具有独特的群落结构和生物多样性特征,研究城市化影响的半自然森林更新特征,可以为城市生物多样性保育与地带性植被恢复重建提供借鉴。文章以上海辰山残存半自然森林1 hm^2固定监测样地为对象,调查分析半自然森林更新苗的物种组成特征。结果表明:1 hm^2样地更新苗共5 937株,分属22科32属37种,其中上海自然分布的乡土种占57.89%,明显高于城市人工林的乡土树种比例;香樟、构树、天竺桂、朴树和小蜡等5种更新苗占总数的57.2%,聚集性分布格局明显;在11个植被群落类型中,各类型更新苗的丰富度普遍较高,不同树种的更新苗存活和发育潜力存在差异。     

瓠瓜(Lagenaria siceraria)白粉病抗性的全基因组关联分析

白粉病是瓜类作物重要病害之一。本研究采用苗期人工接种方法,研究了117份瓠瓜微核心种质对白粉病的抗性,并基于不同材料的病情指数和基因型数据,进行全基因组关联分析(genome-wide association analysis,GWAS)。利用2014年和2018年的抗病表型数据,分别鉴定到22个和13个与白粉病抗性相关的SNPs,其中2个SNPs位点在两年试验中同时被检测到。进一步研究发现不同瓠瓜材料的白粉病抗性与其携带有利等位变异数目成正相关。本研究为克隆瓠瓜抗白粉病相关基因、研究其抗病机制提供科学依据,促进了瓠瓜抗白粉病分子育种。     

翠竹及菲白竹LEA3同源基因的克隆及序列分析

竹类植物在中国有着非常悠久的栽培历史,并被广泛应用于园林、造林、庭院以及食品等行业。本研究以翠竹和菲白竹为材料提取其叶片总DNA,采用PCR方法获得翠竹SpLEA3基因与菲白竹SfLEA3-1、SfLEA3-2基因;其中SpLEA3全长804 bp,编码195个氨基酸,GC含量为68.4%;SfLEA3-1全长803 bp,编码195个氨基酸,GC含量为68.2%;SfLEA3-2全长557 bp,编码144个氨基酸,GC含量为67.8%。通过ProtParamy等生物软件分析,SpLEA3、SfLEA3-1和SfLEA3-2与贵州悬竹LEA3基因同源性高达61%以上,且3个基因编码蛋白均属于亲水性蛋白。源自2个竹种LEA3基因均包含一个完整的开放阅读框,其编码的氨基酸含有4~6个由11个氨基酸组成的保守基元序列。本研究不仅为深入了解竹类植物抗旱的分子机理研究提供了基础数据,也为竹类植物的抗旱育种后续研究提供了科学依据。     

基于高通量测序的露地菊(Chysanthemum×grandiflora)盐胁迫转录组分析

为揭示露地菊生长发育及耐盐胁迫的应答机制和分子基础,本研究以盐胁迫处理的露地菊及其对照为材料,使用Illumina Hiseq2500高通量测序平台对转录组进行测序,分别获得了60370448和71415448条Clean reads,通过序列拼接组装得到45591条Unigene,平均长度724 bp。有37675条Unigene在七大数据库(COG,GO,KEGG,KOG,Pfam,Swiss-Prot,NR)中得到注释。通过比对露地菊盐胁迫处理组和对照组样品间Unigene的表达量及在各数据库中的注释情况,统计得到:有4143条差异表达基因获得注释;有2441条差异表达基因在GO数据库中获得功能注释;注释到COG数据库中的2281条差异表达基因依据功能可分为25类;有1062条差异基因映射到KEGGPathway数据库中,涉及了199个代谢通路,包括核糖体途径、植物激素信号传导途径、淀粉蔗糖代谢、碳代谢、氨基酸的生物合成等。本研究获得的转录组数据将有助于揭示露地菊生长发育及耐盐胁迫的应答机制和分子基础,及相关抗性基因的挖掘和分子辅助育种等方面的研究。     

高粱光敏色素A基因的克隆、蛋白结构与长短日照诱导表达模式

光敏色素是一类红光/远红光受体,对植物光形态建成、外部形态和生理功能等方面起着重要的调节作用。为明晰光敏色素A在调控高粱光形态建成发育过程中的作用,本研究比较分析光温敏感程度不同的高粱品种Btx623、Rio、Tx430和LH645的PhyA基因序列和PHYA蛋白的保守结构域,利用qRT-PCR明确长短日照条件处理下PhyA基因的表达模式。结果表明,Btx623、Rio和Tx430之间PhyA基因序列存在非同义突变,LH645在PhyA基因的第7个外显子上插入了5 bp碱基序列;高粱PhyA蛋白中含有1个PAS-2结构域、1个GAF结构域、1个Phytochrome结构域、2个PAS结构域、1个组氨酸激酶A结构域和1个类似组氨酸激酶的ATP激酶结构;遗传进化树分析表明,单子叶和双子叶植物PhyA蛋白明显聚为2大类,高粱PhyA、谷子PhyA和玉米PhyA1、PhyA2聚为一个亚类,相互间亲缘关系较近,与水稻PhyA同源性相对较远;在短日照(SD)条件下各时期PhyA基因转录水平变化显著,Rio PhyA基因转录水平较Btx623和Tx430变化明显,LH645 PhyA基因转录变化不显著,在长日照(LD)条件下Rio PhyA基因转录水平低于Btx623和Tx430,LH645 PhyA基因转录水平变化不显著。以上结果表明,PhyA受光周期诱导,推测其在高粱光形态建成中起重要调控作用。本研究为进一步解析PhyA基因功能及其在调控高粱光形态建成发育过程中的作用提供理论基础。