TaTPP-7A positively feedback regulates grain filling and wheat grain yield through T6P-SnRK1 signalling pathway and sugar–ABA interaction

Grain size and filling are two key determinants of grain thousand-kernel weight (TKW) and crop yield, therefore they have undergone strong selection since cereal was domesticated. Genetic dissection of the two traits will improve yield potential in crops. A quantitative trait locus significantly associated with wheat grain TKW was detected on chromosome 7AS flanked by a simple sequence repeat marker of Wmc17 in Chinese wheat 262 mini-core collection by genome-wide association study. Combined with the bulked segregant RNA-sequencing (BSR-seq) analysis of an F₂ genetic segregation population with extremely different TKW traits, a candidate trehalose-6-phosphate phosphatase gene located at 135.0 Mb (CS V1.0), designated as TaTPP-7A, was identified. This gene was specifically expressed in developing grains and strongly influenced grain filling and size. Overexpression (OE) of TaTPP-7A in wheat enhanced grain TKW and wheat yield greatly. Detailed analysis revealed that OE of TaTPP-7A significantly increased the expression levels of starch synthesis- and senescence-related genes involved in abscisic acid (ABA) and ethylene pathways. Moreover, most of the sucrose metabolism and starch regulation-related genes were potentially regulated by SnRK1. In addition, TaTPP-7A is a crucial domestication- and breeding-targeted gene and it feedback regulates sucrose lysis, flux, and utilization in the grain endosperm mainly through the T6P-SnRK1 pathway and sugar–ABA interaction. Thus, we confirmed the T6P signalling pathway as the central regulatory system for sucrose allocation and source–sink interactions in wheat grains and propose that the trehalose pathway components have great potential to increase yields in cereal crops.     

大河岸线生态完整性内涵及评估方法——以长江岸线为例

大河岸线作为流域的重要生态空间,其内涵属性、空间格局和生态功能都具有显著的整体性。从岸线的陆域原真性、水陆生态渐变性、生境依存性及生物多样性等维度,分析了大河岸线生态完整性的科学内涵。结合国内外河流岸线生态评价研究,建立了大河岸线生态完整性的科学表征体系,提出各项指标评估和综合分级方法,并对长江岸线生态完整性进行了评估。结果表明,长江岸线生态完整性总体处于中等水平,从上游到下游沿岸各地市岸段生态完整性得分总体逐步降低;中上游地市岸段受到滨江湿地和洲岛发育程度及湿地保护状况等因素制约,下游地市岸段则受土地开发强度、自然岸线保有率、水陆连通性等制约。虽然中下游岸线有较高的开发程度且受长江大堤的约束,生态完整性受到明显影响,但在相同条件下生态完整性仍然有较大差异性,部分地区如铜陵段、安庆段和上海段仍保持了较高的完整性。大河岸线生态完整性的评估有助于形成既尊重自然规律、保护生态环境、保障防洪安全及供水安全,又能支撑社会经济发展、促进高效集约利用的长江生态岸线格局。     

Genome-wide identification,classification, and expression profiling of serine esterases and other esterase-related proteins in the tobacco hornworm,Manduca sexta

Serine esterases (SEs) are hydrolases that catalyze the conversion of carboxylic esters into acids and alcohols. Lipases and carboxylesterases constitute two major groups of SEs. Although over a hundred of insect genomes are known, systematic identification and classification of SEs are rarely performed, likely due to large size and complex composition of the gene family in each species. Considering their key roles in lipid metabolism and other physiological processes, we have categorized 144 M. sexta SEs and SE homologs (SEHs), 114 of which contain a motif of GXSXG. Multiple sequence alignment and phylogenetic tree analysis have revealed 39 neutral lipases (NLs), 3 neutral lipase homologs (NLHs), 11 acidic lipases (ALs), 3 acidic lipase homologs (ALHs), a lipase-3, a triglyceride lipase, a monoglyceride lipase, a hormone-sensitive lipase, and a GDSL lipase. Eighty-three carboxylesterase genes encode 29 α-esterases (AEs), 12 AEHs (e.g., SEH4-1–3), 20 feruloyl esterases (FEs), 2 FEHs, 2 β-esterases (BEs), 2 integument esterases (IEs), 1 IEH, 4 juvenile hormone esterases, 2 acetylcholinesterases, gliotactin, 6 neuroligins, neurotactin, and an uncharacteristic esterase homolog. In addition to these GXSXG proteins, we have identified 26 phospholipases and 13 thioesterases. Expression profiling of these genes in specific tissues and stages has provided insights into their functions including digestion, detoxification, hormone processing, neurotransmission, reproduction, and developmental regulation. In summary, we have established a framework of information on SEs and related proteins in M. sexta to stimulate their research in the model species and comparative investigations in agricultural pests or disease vectors.     

Universal two-dimensional labelled probe-mediated melting curve analysis based on multiplex PCR for rapid typing of Plasmodium in a single closed tube

Currently, malaria is still one of the major public health problems commonly caused by the four Plasmodium species. The similar symptoms of malaria and the COVID-19 epidemic of fever or fatigue lead to frequent misdiagnosis. The disadvantages of existing detection methods, such as time-consuming, costly, complicated operation, need for experienced technicians, and indistinguishable typing, lead to difficulties in meeting the clinical requirements of rapid, easy, and accurate typing of common Plasmodium species. In this study, we developed and optimized a universal two-dimensional labelled probe-mediated melting curve analysis (UP-MCA) assay based on multiplex and asymmetric PCR for rapid and accurate typing of five Plasmodium species, including novel human Plasmodium, Plasmodium knowlesi (Pk), in a single closed tube following genome extraction. The assay showed a limit of detection (LOD) of 10 copies per reaction and could accurately distinguish Plasmodium species from intra-plasmodium and other pathogens. Additionally, we proposed and validated different methods of fluorescence quenching and tag design for probes that are suitable for UP-MCA assays. Moreover, the clinical performance of the Plasmodium UP-MCA assay using a base-quenched universal probe was evaluated using 226 samples and showed a sensitivity of 100% (164/164) and specificity of 100% (62/62) at a 99% confidence interval, with the microscopy method as the gold standard. In summary, the UP-MCA assay showed excellent sensitivity, specificity, and accuracy for genotyping Plasmodium species spp. Additionally, it facilitates convenient and rapid Plasmodium detection in routine clinical practice and has great potential for clinical translation.     

四川黑竹沟地区非飞行小型兽类物种多样性与群落组成

四川黑竹沟国家级自然保护区位于生物多样性丰富的凉山山系, 在保护和维持区域生物多样性方面具有极其重要的地位, 然而对该自然保护区兽类群落的研究却极为缺乏。为了了解该自然保护区及周边的小型兽类群落,本文作者在2018年4-10月调查了该区域的非飞行小型兽类物种多样性及其群落组成。在海拔1,537-3,830 m间共设置样方184个, 布设鼠夹9,016铗日, 捕获小型兽类536只, 隶属4目7科13属21种。包括滇攀鼠(Vernaya fulva)和等齿鼩鼹(Uropsilus aequodonenia)两个稀有物种在内的共计9个物种为该地区首次报道, 丰富了物种分布记录。结合历史资料, 黑竹沟地区共记录小型兽类43种, 隶属4目9科28属。在43种小型兽类中, 东洋界物种37种, 占绝对优势(86%), 分布型以喜马拉雅-横断山型占优, 为18种(占东洋界48.6%)。本次调查捕获的21种小型兽类中, 群落优势种为中华姬鼠(Apodemus draco, 33.2%)、北社鼠(Niviventer confucianus, 21.3%)和中华绒鼠(Eothenomys chinensis, 12.7%)。随着海拔上升, 群落优势种组成发生变化, 由北社鼠 + 中华姬鼠 + 针毛鼠(Niviventer fulvescens) + 短尾鼩(Anourosorex squamipes)群落, 转变为中华绒鼠 + 中华姬鼠 + 凉山沟牙田鼠(Proedromys liangshanensis) + 西南绒鼠(Eothenomys custos)群落。群落物种区系的分布型组成随着海拔升高而发生变化, 喜马拉雅-横断山型物种随海拔升高所占比例增大; 分布于喜马拉雅-横断山的特有种分布的下限、中点和上限平均海拔都高于非特有种, 且特有种和非特有种的中点和下限平均海拔差异显著(n = 21, df = 19, P = 0.013; n = 21, df = 19, P < 0.01), 说明该区域物种具有明显的东洋界特征, 中高海拔主要由特有种构成。本研究丰富了黑竹沟地区小型兽类及群落多样性的数据资料, 有利于该地区物种多样性的研究和保护。     

Meeting report on the 2019 international symposium of molecular design breeding in animals (Yangling,China) with the consensus on genome-editing agricultural animals and their regulation

Transgenic Research -     

刺梨GL2同源基因的克隆、系谱树和表达分析

为了观察刺梨果实的果刺细胞学发育过程,该研究以刺梨‘贵农5号''的cDNA为模板,通过RACE克隆获得刺梨中与拟南芥表皮毛形成GL2的同源基因RrGL2,并对该基因进行生物信息学分析和表达分析。结果表明:(1)刺结构在花芽形成早期基部内的细胞首先不断分裂,向外继续发育,中部的细胞变细、变长形成“针”状结构,顶部的细胞逐渐木质化使刺变硬,形成果刺。(2)通过RACE扩增得到RrGL2的cDNA全长2 292 bp,编码763 aa氨基酸。(3)RrGL2具有Homeodomain同源结构域和StAR磷脂酰胆碱转移蛋白的结构域,RrGL2与其他物种编码的GL2氨基酸同源性高度相似,并且系谱树分析揭示刺梨RrGL2和野草莓的GL2密切相关。(4)qRT-PCR分析表明,RrGL2在茎和果实中的表达水平高于其他组织,在花后7周果刺中的表达最高,是3周和5周果刺中的7.87倍和2.10倍。综上结果发现RrGL2的功能与果刺的形成发育密切相关,该研究为刺梨中刺形成的分子机制和育种提供了理论基础。     

蒙古沙冬青群落区系组成及分类研究

蒙古沙冬青是西北地区珍稀濒危树种,探究区系组成和群落特征对于维持西北干旱荒漠区脆弱生态系统具有重要意义。该研究调查了26个蒙古沙冬青群落样地,采用TWINSPAN进行群落分类,同时利用典范对应分析(CCA)分析沙冬青群落物种组成与生态因子的关系,为珍稀濒危植物蒙古沙冬青的保护提供基础数据。结果表明:(1)蒙古沙冬青群落物种组成以豆科、禾本科、菊科和藜科植物为主,隶属于28科63属90种,区系组成以地中海、北温带分布及世界分布为主。(2)TWINSPAN等级分类可将26个蒙古沙冬青样地划分为9个群落类型,即蒙古沙冬青+蓍状亚菊群落、蒙古沙冬青+红砂群落、蒙古沙冬青+新巴黄耆群落、蒙古沙冬青+无芒隐子草群落、蒙古沙冬青+霸王群落、蒙古沙冬青+驼绒藜群落、蒙古沙冬青+白刺群落、蒙古沙冬青+沙鞭群落和蒙古沙冬青+毛刺锦鸡儿群落。(3)CCA排序分析显示,年均降雨量(F=2.8,P=0.002)、生长季降雨量(F=2.6,P=0.002)、速效氮(F=2.1,P=0.006)、太阳辐射量(F=2.1,P=0.008)、蒸汽压(F=1.9,P=0.006)、土壤有机质(F=1.7,P=0.04)、纬度(F=1.7,P=0.006)是影响蒙古沙冬青群落组成的主要生态因子。     

2种调查方法对四川黑竹沟国家级自然保护区3种雉类种群密度调查的比较

种群数量是物种的重要生态学基础资料,合适的密度调查方法是数量估算的基础。2016年4-5月,采用广泛应用于鸡形目Galliformes鸟类种群密度调查的样线法和样点法,调查了四川黑竹沟国家级自然保护区3种鸡形目鸟类(白腹锦鸡Chrysolophus amherstiae、红腹角雉Tragopan temminckii和血雉Ithaginis cruentus)的种群密度。样线法和样点法估算的雄体密度分别是:白腹锦鸡1.20只/km^2和(6.31±0.98)只/km^2,红腹角雉5.41只/km^2和(0.39±0.17)只/km^2,血雉3.01只/km^2和(5.97±2.70)只/km^2。除红腹角雉外,样点法估算的白腹锦鸡、血雉种群密度均大于样线法。建议针对不同鸡形目鸟类采用不同的调查方法,并尽量扩大样本数量,从而提高调查结果的准确性。