水稻CSC11介导干热风/干旱诱导的钙信号调控雄蕊发育

【目的】水稻花期偶遇干热风/干旱,导致脆弱的生殖细胞快速失水,极大地降低产量,这一过程中钙离子作为通用的第二信使传导了干旱或其他逆境信号,但背后的分子机制尚不清楚。分析钙离子透过性胁迫反应阳离子通道家族（calcium-permeable stress-responsive cation channels,CSCs）基因的生理和分子功能,为研究作物干热风的感应机制提供新的理论基础和思路。【方法】采用电生理学和遗传学方法,利用双电极电压钳技术在水稻中鉴定得到一个具有典型特征的受体类-钙通道蛋白,名为OsCSC11,对其蛋白序列进行生物信息学和进化关系分析。运用qRT-PCR和GUS报告基因活性分析确认OsCSC11的表达模式,在拟南芥原生质体细胞和洋葱表皮细胞中瞬时表达OsCSC11-GFP融合蛋白,验证OsCSC11的亚细胞定位;同时利用CRISPR/Cas9基因编辑技术获得OsCSC11的突变体,并通过细胞学等手段分析突变体表型和相关生理功能。【结果】蛋白序列比对发现,OsCSC11具有CSCs家族成员典型的保守结构域DUF221,但与其他成员序列差异大,存在不同于其他成员的特异结构域（motif）属于独立的亚家族。OsCSC11主要在水稻的花药和叶片中表达,进一步分析发现全长OsCSC11处于静息状态,可被高渗透溶液激活;但是删除N端156氨基酸（TM1-3）之后的OsCSC11^ΔTM1-3具有组成型的通道活性,特异选择钙、镁二价阳离子;推测TM1-3是这类通道的受体结构域,感应干热风胁迫,而OsCSC11^ΔTM1-3区域负责钙信号产生。OsCSC11和OsCSC11^ΔTM1-3均定位在细胞质膜上,与其干热风的受体功能相适应。与野生型相比,功能缺失突变体oscsc11-1和oscsc11-2的雄蕊较小、花药表面蹙皱,整体多呈弯曲状态,花粉含水量较低,败育率高达60%—70%。【结论】OsCSC11是水稻感应短期干热风/干旱刺激、介导钙离子内流,调控花药水分状态和花粉发育的受体类钙通道,可能参与了水稻雄蕊应对干热风的原初感应过程。     

龟纹瓢虫生殖系统及卵子发生观察

【目的】龟纹瓢虫捕食蚜虫、粉虱、木虱等多种害虫,是农林业重要天敌昆虫。掌握龟纹瓢虫雌雄内生殖系统结构及卵细胞生成过程,为龟纹瓢虫人工繁育提供理论依据。【方法】采用组织石蜡切片技术和内部解剖,在光学显微镜下观察龟纹瓢虫雌雄成虫内生殖器及卵子发生过程。【结果】龟纹瓢虫雄性内生殖系统包括1对精巢、1对雄性附腺、1对输精管、贮精囊、射精管;雌性内生殖系统包括1对卵巢、1对侧输卵管、中输卵管、受精囊、生殖腔,每侧卵巢含11~13根卵巢管;卵巢管生长区可见4粒发育程度不同的卵母细胞,根据卵黄沉淀情况、滤泡细胞形态变化将卵子发生划分为前期、中期、后期、末期4个时期。【结论】卵巢结构、卵巢管和卵母细胞数量有助于解释龟纹瓢虫产卵量,不适宜的饲养条件可能抑制卵巢发育,进而导致龟纹瓢虫产卵量发生变化。因此,为实现龟纹瓢虫的规模化应用,人工繁育中应创造最优营养和环境条件以保障产生更多成熟卵细胞。     

Thermal shock induction of triploidy in Atlantic cod (Gadus morhua L.)

The effects of thermal treatments on induction of triploidy in Atlantic cod have been investigated. Cold shock [−1.7±0.1°C at 20 min post fertilization (PF) for 2 h] was based on a previously developed protocol, and heat shocks, below the lethal threshold of 24°C, were at 16, 18 or 20°C applied 20, 30 or 40 min PF for 20 min. Cold shock did not affect larval survival and was ineffective for producing triploids (range 0–4%). A heat shock of 20°C at 20 min PF generated the highest percentages (range 66–100%) of triploid larvae at hatching, with survival ranging from 10% to 20% relative to the controls. Lower heat shock temperatures or delayed shocks increased survival but decreased the number of triploids, providing no net gain in triploid yield (range 1–9%). Heat shocks applied later than 20 min PF produced 2–4% tetraploid larvae at hatching. A thermal shock of 20°C initiated at 20 min PF and lasting 20 min proved to be the most generally efficient treatment for induction of triploidy in Atlantic cod.     

不同断奶日龄对奶公犊生长性能和体尺指标的影响

30头新生荷斯坦奶公犊出生日期和体重相近,随机分成6组,对照组按常规饲喂鲜牛奶,60日龄断奶;各试验组分别于10、20、30、40和50日龄断奶,试验期为60d。结果表明:试验结束时,20日龄断奶组、30日龄断奶组的体重、平均日增重均显著高于对照组,40日龄断奶组、50日龄断奶组与对照组间无明显差异;各试验组犊牛与对照组在体长和体斜长上无显著差异。     

Changes in estrogen receptor expression in the chick thymus during late embryonic development

In chickens, although estrogen receptors (ER) are reported to be associated with the immunological processes, detailed information about the differences in ER expression in the tissues related to the development of lymphocytes is not fully known, especially during the developmental stage. To learn more about this immunological relationship, we used semi‐quantitative polymerase chain reaction method to detect the ER expression levels in the thymus tissues of chicks during the developmental stage. Furthermore, ER‐expressing cells were detected by immunohistochemistry. The results of this study show that the expression level of ER increased on embryonic day 16 and decreased on day 20. Furthermore, ER expression was significantly higher in male than in female chickens at day 16. The increased expression on day 16 and decreased level on day 20 were also reproduced in the incidence of immunoreactive cells, although there was a 1‐day delay in the elevated incidence of the cells. This study revealed the changes in ER expression and the incidence of ER‐positive cells in the thymus of chickens during the developmental stage.     

Population consequences of behaviour in the European bitterling (Rhodeus sericeus Cyprinidae)

Abstract –  The European bitterling is a freshwater fish with an unusual spawning symbiosis with freshwater mussels. Female bitterling possess long ovipositors that they use to place their eggs onto the gills of a mussel. Males fertilise the eggs by releasing sperm into the inhalant siphon of the mussel. The embryos develop inside the mussel for approximately a month, eventually leaving the mussel as actively swimming larvae. Because they use a discrete spawning site that can be readily manipulated they represent an ideal model for linking reproductive decisions with population dynamics. Bitterling have been used in field and large-scale pool experiments, in combination with modelling and population and genetic studies to investigate the population consequences of behaviour. Here we show how male mating tactics, female mate and oviposition decisions, and aggressive interference competition among juveniles, may have significant impacts on bitterling population size and population genetics.     

褐飞虱杀雄菌属共生菌Arsenophonus的传递方式与生物学意义初探

通过阳性(含Arsenophonus)与阴性(不含Arsenophonus)褐飞虱初羽化成虫的正反交试验,发现子代感染Arsenophonus的情况由其母本决定,而与父本无关,表明褐飞虱体内的Arsenophonus可由母体垂直传递给子代。初羽化阳性和阴性成虫的配对实验结果表明,Arsenophonus对褐飞虱后代数量及性比没有显著的作用,无雄性致死作用。通过黄绿绿僵菌ARSEF1764菌株对初羽化雌成虫褐飞虱的接种实验,发现阴性试虫的感病率为55.2%,显著高于阳性试虫的32.5%,表明感染Arsenophonus有助于提高褐飞虱对黄绿绿僵菌ARSEF1764菌株的抵抗能力。     

温敏雄性不育小麦BNS366花粉败育的细胞学观察

为了明确温敏雄性不育小麦BNS366雄性败育的细胞学机理,采用醋酸洋红染色法观察减数分裂和小孢子发育过程,用1%I2-KI溶液染色进行花粉育性统计;以温敏不育系BNS和常规品种扬麦13作为对照。结果表明,BNS366的减数分裂和小孢子发育过程与BNS完全一致,即:在减数分裂过程中,中期Ⅰ染色体出现滞后现象,在形成二分体和四分体时细胞质分裂不均匀,出现三孢现象;小孢子发育过程中,只观察到单核靠边期,最后核物质解体出现空孢现象。而扬麦13在减数分裂过程中染色体表现正常,小孢子发育过程中,观察到单核期→双核期→三核期。开花期,BNS366和BNS花粉均表现典败和圆败,扬麦13可育。减数分裂表现出的异常行为是导致BNS366花粉败育的原因之一,单核靠边期是其败育的关键时期。     

5种细胞质雄性不育小麦败育过程中活性氧代谢保护酶的响应

为了解异质雄性不育小麦败育过程中活性氧代谢中几种保护酶的变化特点,以5种小麦雄性不育系U116A、Ju116A、Va116A、C6116A、K116A及其相应保持系116B为材料,分析了小麦雄性不育系在花药不同发育时期的过氧化物酶(POD)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)活性与保持系的差异。结果表明,不育花药的POD活性自单核早期后均比同一发育时期的可育花药高;自单核晚期开始,不育系CAT活性总体上呈下降趋势,而可育系则呈升高趋势;受试材料花药中SOD活性总体呈上升趋势,且不育系高于保持系。经差异分析,不育系U116A、Ju116A、Va116A、C6116A的败育关键期是单核晚期,而K116A的败育关键期则是单核晚期至二核期。以上结果说明,5种异质小麦雄性败育时期有所不同,并且雄性育性与POD、CAT和SOD活性密切相关。     

Development ofNew InDel Marker to Detect Genotypes of Conferring Fertility Restoration of BT-Type Cytoplasmic Male Sterility inRice简

Restorer line breeding is an important approach to enhance the heterosis and improve the yields of japonica hybrid rice. To improve the selection efficiency of restorer lines for BT-type cytoplasmic male sterility (CMS) in japonica rice, a functional marker InDel-Rf-1a based on the difference of nucleotide sequence in Rf-1a locus between BT-type CMS lines and restorer lines was developed to detect the genotypes of different rice materials. Conventional indica rice varieties, restorer and maintainer lines without 574 bp deletion could restore the fertility for BT-type CMS in japonica rice. By contrast, most conventional japonica rice varieties except Aichi 106 and Yijing 12, with genotype of rf-1arf-1a showed the 574 bp deletion maintained sterility for BT-type CMS lines. To further verify the effect of genotyping detection in Rf-1a locus, this marker was also used to amplify the genomic DNA in different japonica rice restorer lines, CMS lines, hybrids and F₂ segregation population, and three genotypes in Rf-1a locus could be distinguished distinctly. Therefore, the marker InDel-Rf-1a could be widely used for genetic identification and marker-assisted selection (MAS) in breeding japonica restorer lines.