早熟、高产转基因抗虫常规棉品种——湘K25

概述了湘K25的选育经过，介绍了湘K25的特征特性、产量、纤维品质和抗病、抗虫等特点，并总结了其关键栽培技术。     

优质、高产转基因抗虫棉杂交种苏杂668的选育与栽培技术

本文概述了苏杂668的选育过程，阐明了苏杂668的特征特性、产量水平、纤维品质和抗性表现，并提出其高产栽培技术措施。     

Artificial fertilization and generating families for a selective breeding programme of large yellow croaker ( Larimichthys crocea )

Large yellow croaker is an important marine aquaculture species in China. The aim was to determine an appropriate protocol of artificial fertilization for family construction in the breeding programme based on two trials. In trial 1, luteinizing hormone-releasing hormone A3 (LHRHA3) was injected once, with a dosage of 2 μg/kg for females and 1 μg/kg for males. The latency time was in the range of 30–34 h. The maturation stage was checked by extracting a few eggs with a Pasteur pipette. The fertilization rate and hatching rate were 27 and 52%, respectively. The percentage of females with spawning difficulties was 30%. In trial 2, the females were injected LHRHA3 twice: with a first dose of 0.8 μg/kg and a second dose of 2 μg/kg, at an interval of 10 h, whereas the males were still injected once. The latency time was in the range of 29.5–35 h, determined by only observing courtship behaviour of males. The females with spawning difficulties decreased to 10%, and the fertilization rate and hatching rate also improved to 41 and 62%, respectively.     

Two biomass preparation methods provide insights into studying microbial communities of intestinal mucosa in grass carp (Ctenopharyngodon idellus)

Animal digestive tract is habitat for a large number of autochthonous microbiota, which play central roles in multiple biological and physiological processes of the host. In this study, two different micro‐biomass preparation methods were employed to evaluate the diversity of intestinal mucosa‐associated microbiota in grass carp (Ctenopharyngodon idellus). Genomic DNAs were isolated either directly from intestinal mucosal samples (group A), or from micro‐biomass after microbial dissociation (group B). Community richness, diversity and evenness indices were all higher in group B, but differences were not statistically significant (P = 0.97, P = 0.33, P = 0.34 respectively). Furthermore, group B samples exhibited an increased ratio of bacterial DNA in comparison with group A samples, but the difference was also not statistically significant (P = 0.74). In addition, there were no statistically significant differences between the two groups (P > 0.05) at the taxonomic level. Our results support previous findings that there exists a great abundance of the intestinal mucosa‐adherent microbiota in the grass carp; among these, Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Spirochaetes and Fusobacteria were the most common phyla. Within these microbiota, Paenibacillus, Bacteroides, Bacillus and Cetobacterium genera comprise the majority of the community, implicating their functional importance (e.g. as probiotics) to their host. Our results contribute towards a better understanding of the intestinal microbial profile of grass carp. Both micro‐biomass preparation techniques proved to be feasible for studying mucosa‐adherent microbiota of grass carp; however, the second method (group B) provides a protocol that is somewhat more effective than the first method (group A).     

The influence of diet on the grass carp intestinal microbiota and bile acids

To elucidate the influence of different diet on the intestinal microbe and bile acids, we characterized the microbiota and bile acids in the hindgut content of grass carp fed on formula feed (FF group) or Sudan grass (SG group). Fusobacteria and Proteobacteria were significantly more represented in FF group than in SG group whereas Bacteroidetes was significantly more abundant in SG group than in FF group. Simpson diversity was significantly higher in FF group than in SG group (t = 2.33, P < 0.05). Chenodeoxycholic acid (CDCA) was the most abundant primary bile acid in the two groups, with average concentrations of 1.03 ± 0.62 and 4.44 ± 1.80 ng mg^?1 in SG and FF group respectively. The most abundant secondary bile acid was deoxycholic acid (DCA) in SG group and ursodeoxycholic acid (UDCA) in FF group, with average concentrations of 0.17 ± 0.06 and 2.67 ± 0.88 ng mg^?1 respectively. UDCA is significantly more abundant in FF group than in SG group, and the total bile acids were higher in FF group than in SG group. Cetobacterium and Fusobacteriaceae U114 were significantly related with the concentrations of CDCA (r = 0.85, P < 0.05 and r = 0.82, P < 0.05 respectively) and UDCA (r = 0.92, P < 0.01 and r = 0.92, P < 0.01 respectively). However, Bacteroides was negatively related with the concentration of UDCA (r = ?0.67, P < 0.05). Overall, there existed certain relationship between the intestinal microbes and the faecal bile acids, and they were both influenced by the diet.     

Impacts of diet on hindgut microbiota and short‐chain fatty acids in grass carp (Ctenopharyngodon idellus)

Diet is known to influence intestinal microbiota in fish, but the specifics of these impacts are still poorly understood. Different protein/fibre ratio diets may result in differing structures and activities of gut microbiota. We examined the hindgut microbiome of grass carp (Ctenopharyngodon idellus) fed three different diets: fish meal (FM, high protein – low fibre), Sudan grass (SG, high fibre – low protein) and compound feed (CF, intermediate). Microbial profiles of fish fed on FM were significantly different from profiles of fish fed CF and SG (F = 18.85, p < .01). Cetobacterium, known to be positively associated with protein digestion, was the dominant microbial group in FM samples (approximately 75.7%), while Lachnospiraceae and Erysipelotrichaceae, thought to be involved in fermentation of plant polysaccharides, were dominant in CF and SG samples (46.8% and 42.9% respectively). Network analyses indicated that the abundance of Lachnospiraceae and Erysipelotrichaceae was in a significantly positive correlation (r = .895, p = .001). Short‐chain fatty acid (SCFA) levels may indicate that the digestibility of diet by microbiota in the grass carp gut decreased from FM to SG (FM>CF>SG). Overall low SCFA levels indicate that hindgut fermentation probably provides a low proportion of energy requirements in grass carp.     

花生栽野种间杂交异源多倍化早期世代基因表达变化分析

为了进一步认识花生种间杂交异源多倍体进化过程中的基因表达变化规律,采用cDNA-HFO-TAG技术,研究花生种间杂交组合(四倍体栽培种‘仲恺花4号’×二倍体野生种Arachis. diogio)杂种F_1和早期多倍体世代S0~S3的基因表达变化情况。14条HFO-TAG引物共扩增出121条cDNA片段,其中差异片段84个,主要包括三种类型:亲本转录物完全沉默(3个),双亲转录物在后代部分材料中沉默(59个)和新转录物激活(22个),上述变化在F1代即开始发生。筛选其中大小为500~2 000 bp的35个TDFs进行克隆测序,有27个和NCBI数据库中已录入的基因具有较高的相似性,包括抗逆相关基因(10条)、未知功能蛋白基因(8条)、能量与代谢相关基因(7条)和转录因子相关的基因(2条)。这些研究结果进一步表明在花生种间杂交异源多倍化早期发生着快速、剧烈的基因表达变化,从中获得的差异基因片段,有助于了解花生属种间杂交异源多倍化早期分子机制变化,这对有效利用野生花生种质优异基因具有重要意义。cDNA-HFO-TAG技术简单、有效且实用,完全适用于花生属基因表达变化研究,可以作为花生属及其它物种基因表达变化分析技术的有效补充。     

基于Meta分析评价秸秆覆盖对我国北方半干旱区不同生态区域小麦产量的影响

为了明确秸秆覆盖对我国北方半干旱区不同降雨量和积温条件下小麦产量的影响,利用Meta定量分析方法分析秸秆覆盖和不同秸秆覆盖量对我国北方半干旱区不同降雨量和积温条件下小麦产量的影响。以“半干旱区”、“秸秆覆盖”和“小麦产量”为关键词搜集、筛选并整理1970-2019年公开发表的相关文献,建立综合文献数据库。分析秸秆覆盖、不同秸秆覆盖量、不同年均降雨量和不同年均积温与小麦产量的关系。结果表明,秸秆覆盖可以增加北方半干旱区不同降雨量和积温区域的小麦产量,主要通过增加有效穗数使小麦平均增产12.77%;当秸秆覆盖量为6 000~9 000kg/hm ²时,小麦增产率最高,为14.49%（9.01%~20.52%,CI>95%）;当年均降雨量为200~400mm时,小麦增产率最高,为19.36%（9.86%~28.49%,CI>95%）;当年均积温为0℃~3 000℃时,小麦增产率最高,为19.56%（16.36%~22.39%,CI>95%）。此研究结果可为我国北方半干旱区不同降雨量和积温区域采用秸秆覆盖技术栽培小麦提供理论依据。     

EMS诱导水稻‘一目惚’表型变异和部分基因表达变异的比较

本研究利用甲基磺酸乙酯(Ethy methan sulfonate)试剂胁迫诱导粳稻品种‘一目惚’(Hitomebore),获得性状稳定遗传的水稻矮杆突变体(E58),对株高、分蘖、千粒重等表现与相同环境生长下的野生型‘一目惚’对照(E63)进行对比分析,结果表明,该突变体田间表现为植株矮化,分蘖增多,千粒重减少,且株高与对照相比呈极显著差异,分蘖呈显著差异,千粒重呈极显著差异。EMS诱导的序列变异可能对基因表达产生影响,进而引起水稻矮化的表型变异。根据前人试验结果,应用Real-time PCR对与胁迫及矮化相关的39对引物中筛选出24对引物进行Real-Time PCR扩增,结果表明:引物OsNR1、OsP5CS、OsGS1;1、OsGDH1、OsHKT1;5、OsNRT1;1、OsGDH3、OsAKT1、OsAMT2;3、OsGS1;2、OsSOS1、OsNHX2、OsNRT2;1、OHAK16、OsGS2、OsHAK1、OsGDH2、OsAMT3;3在E63、E58表达差异极显著;OsHAK10、OsNHX1、OsAMT1;1表达差异显著;OsNADHGOGAT、OsFd-GOGAT、OsGS1;3表达差异不显著。EMS诱导对部分水稻基因表达产生影响,从而影响水稻表现型,为进一步工作提供科学依据。     

一个潜在催化莱鲍迪D苷合成的新型糖基转移酶

尿苷二磷酸糖基转移酶(uridine diphosphate glycosyltransferases,UGTs)催化糖基转移反应,与植物次生代谢密切相关。本研究根据甜叶菊(Stevia rebaudiana)转录组数据库,克隆到一个催化莱鲍迪D苷(rebaudioside D,RD)合成的新型糖基转移酶候选基因,对其开展生物信息学分析。结果表明,该基因开放阅读框长1380 bp,编码459个氨基酸,等电点(pI)预测为5.54,理论分子量约49.66 kD,系统发育分析表明该基因与向日葵中的UGT89A2同源,故将其命名为SrUGT89A2。构建pET28a-SrUGT89A2原核表达载体,并在大肠杆菌(BL21(DE3))中诱导表达得到重组蛋白,HPLC检测表明粗酶液能催化甜叶菊提取液形成一个新的色谱峰,该峰保留时间与莱鲍迪D苷一致。经进一步纯化UGT89A2蛋白,添加不同甜菊糖苷标准品为催化底物,但未鉴定出该蛋白催化的具体糖苷。该潜在催化甜菊糖RD苷合成的新型糖基转移酶基因SrUGT89A2的发现,为RD苷的生物合成和甜菊糖苷的生物途径研究提供新的理论依据。