调控动物性状相关长链非编码RNA的研究进展

长链非编码RNA(long non-coding RNA,lncRNA)是基因转录过程中产生的一类长度大于200个核苷酸(nt)的非编码RNA(non-coding RNA,ncRNA)。lncRNA的表达水平通常低于mRNA,且无高度保守序列,缺少开放阅读框,但它们具有更强的组织特异性表达模式。lncRNA可以通过与DNA、RNA(mRNA,miRNA,环状RNA)和蛋白质进行相互作用来发挥其功能,因此可作为信号分子、诱导物等来调节复杂的基因表达网络。作为一种新的调节分子,lncRNA正在成为基因表达调控中新的重要参与者,且近年研究表明,其与家畜动物性状调控密切相连。本文对lncRNA在动物肌肉生长分化、脂肪沉积、毛囊发育和繁殖方面进行了综述,旨在为lncRNA在家畜遗传育种上的应用提供依据。     

胚胎干细胞的研究进展及应用前景

干细胞(Stem Cell)是一类具有分化潜能和自我复制的早期未分化细胞。胚胎干细胞(Embryonic stem cells,ES细胞)是一种早期胚胎内细胞团(inner cell mass,ICM)或原始生殖细胞(primordial germ cell,PGC)经体外分化抑制培养,分离和克隆得到的具有发育全能性的高度未分化细胞。本文综述了胚胎干细胞的形态学、生长特性、免疫学鉴定方法,ES细胞抑制分化和诱导分化的机理以及饲养层、血清和细胞因子等影响胚胎干细胞分离克隆的因素,并进一步阐述了胚胎干细胞的应用前景以及存在的问题。     

Gonadal morphogenesis and sex differentiation in cultured chub mackerel,Scomber japonicus

The timing of primordial germ‐cell (PGC) migration with regard to the gonadal anlagen, gonad formation and sex differentiation was examined histologically in the chub mackerel (Scomber japonicus) at 5–190 days post hatching (dph). At 5 dph, PGCs appeared on the peritoneal epithelium surface or in the mesentery, on the dorsal side of the abdominal cavity. By 10 dph, stromal cells around the PGCs proliferated. The gonadal primordium was formed by 15 dph. The gonadosomatic index was 0.01% at 30 dph and increased thereafter (0.32% in females and 0.04% in males at 160 dph). Ovarian differentiation occurred at 30–40 dph, indicated by ovarian cavity formation (elongation and fusion of the upper and lower ovarian edges). Meiosis was subsequently initiated. A few meiotic oocytes surrounded the cavity at 50 dph; most were in the perinucleolus stage at 60 dph and attained a diameter of 60–70 μm at 190 dph. Testicular differentiation occurred at 30 dph, indicated by the formation of the sperm duct primordium. Spermatogonia gradually proliferated, developing into spermatocytes at the chromatin–nucleolus stage (after 90 dph) and subsequently into spermatids and spermatozoa (160 dph). These data could aid the development of seeding and cell‐engineering technologies for scombrid fish.     

Loss of genetic variability in the captive stocks of tambaqui,Colossoma macropomum (Cuvier, 1818), at breeding centres in Brazil,and their divergence from wild populations

The loss of variability in farmed populations and the risks of interactions with wild populations support the need for the genetic monitoring of species farmed throughout the world. In Brazil, the tambaqui is the most widely farmed native fish species. Despite this, there are no data on the pedigree of the farmed stocks, and the potential for interactions with wild populations in the Amazon basin has raised concerns with regard to the genetic variability of these stocks. The present study analysed sequences of the mitochondrial Control Region and 12 microsatellites to characterize the genetic variability of seven historically important commercial tambaqui breeding centres located in four different regions of Brazil, and compared these sequences with those obtained from individuals collected from a wild population. High levels of genetic diversity were found in the wild population, whereas genetic diversity was reduced in both markers in most captive populations, except for the broodstock located near the Amazon River. High F_ST and D_EST indices were recorded between the wild population and most of the captive stocks analysed. The drastic reduction in genetic diversity found in most captive stocks and the difference between these stocks and the wild population may have been the result of the small size of the founding populations and the absence of breeding management. The renewal of the broodstocks and the application of breeding management techniques are recommended. In the Amazon region, in addition, the use of broodstocks that are genetically very different from local wild populations should be avoided.     

西江流域卷口鱼线粒体D-loop序列的遗传多样性分析

为研究西江流域广西境内卷口鱼(Ptychidio jordani)种群的遗传变异情况,自广西境内6个江段采集了139尾样本,采用PCR与DNA测序技术分析其线粒体D-loop序列的遗传多样性及群体历史动态;139条D-loop序列长度均为725 bp,碱基组成A+T (65.7%)远远高于C+G (34.3%),共检测到变异位点25个,转颠换比R值为11.5。139尾样本共定义23个单倍型。单倍型的NJ系统树以及网络结构图显示,23个单倍型间有2个明显分支,不同地理群体来源的单倍型混杂分布在2个分支中,未能观察到明显的地理聚群。6个群体遗传多样性较好,单倍型多样性Hd=0.71585~0.92063,核苷酸多样性Pi=0.00173~0.00668,遗传分化极其显著(遗传分化系数Fst=0.36737,P<0.001)。AMOVA分析表明,群体内变异占63.26%,群体间为36.74%。中性检验(Tajima’s D= –0.50322,P=0.34600;Fu’s Fs= –5.05210,P=0.08800)与核苷酸错配分布表明,西江流域卷口鱼种群近期内未经历过种群扩张。综上所述,西江流域广西境内的卷口鱼遗传多样性表现为高单倍型、低核苷酸多样性的特征,群体间不同程度的遗传分化表明水坝阻隔及捕捞因素可能促进其发生,而水利梯级开发可能是促进卷口鱼群体遗传分化的首要原因。     

苹果梨花芽分化期核酸代谢规律的研究

采用生化测定的方法对苹果梨花芽分化的核酸代谢进行了研究，结果表明：花芽生理分化期核酸总量高，RNA高于DNA，RNA／DNA比值高，其核酸代谢模式为：RNA双峰，第1峰大于第2峰，DNA单峰，在RNA双峰之间，RNA／DNA呈双峰，蜂值出现时间与RNA双峰相同，第1峰远大于第2峰；花芽形态分化过程中，RNA和DNA变化动态相似，一直处于较高水平且呈增加趋势，RNA高于DNA，雄蕊和紫蕊分化期含量最高，形态分化结束时降至最低。     

薄壳山核桃雄花花芽分化的解剖学研究

为掌握薄壳山核桃(Carya illinoinensis)雄花花芽分化规律,采用石蜡切片方法从显微水平上对浙江建德薄壳山核桃花芽发育过程各阶段特征进行观察,结果表明:3月中下旬分化初期,雄花芽与叶芽无区别;3月下旬鳞片分化期,花序基部逐渐形成突起,进入鳞片化阶段;4月上旬苞片分化期,雄花序自上而下形成弯月形苞片原基,花序顶部呈馒头状;4月中旬进入雄花原基分化期,由苞片基部逐渐形成突起状雄花原基;4月下旬雄蕊原基出现,形成雄蕊原基,产生花药;4月下旬至5月上旬花药分化形成期,花药内形成花粉囊,花粉散出,花序凋谢.     

共轭亚油酸对C2C12肌细胞生脂和生肌分化的影响

本研究分析了共轭亚油酸(CLA)对C2C12肌细胞生脂转分化和生肌分化的影响。分别培养并诱导C2C12鼠源肌细胞生脂转分化和正常的生肌分化,同时分别使用终浓度为50μmol/L的c9,t11-CLA和t10,c12-CLA处理细胞,并设对照组,取生脂转分化第10天和生肌分化第8天的细胞用于实时定量PCR检测,观察c9,t11-CLA和t10,c12-CLA对C2C12肌细胞不同分化的影响。结果表明:1)与对照组相比,c9,t11-CLA促进了C2C12肌细胞的生脂转分化,显著增加了细胞内甘油三酯(TG)含量(P0.05),显著上调了细胞内脂肪酸合成酶(FAS)、CCAAT增强子结合蛋白α(C/EBPα)、过氧化物酶体增殖剂激活受体γ(PPARγ)和脂肪酸结合蛋白4(FABP4)基因的表达水平(P0.05);与对照组相比,t10,c12-CLA则抑制了C2C12肌细胞的生脂转分化,显著减少了细胞内TG含量(P0.05),显著下调了细胞内C/EBPα、PPARγ和FA BP4基因的表达水平(P0.05)。免疫印迹杂交结果显示FAS和FABP4的蛋白质表达水平也发生了与基因表达相一致的变化。2)与对照组相比,t10,c12-CLA抑制了C2C12肌细胞的生肌分化,显著减少了细胞内肌管数/细胞数(P0.05),显著下调了细胞内肌细胞生成素(MYOG)和成肌分化抗原(MYOD)基因的表达水平(P0.05);与对照组相比,c9,t11-CLA则显著上调了细胞内MYOG基因的表达水平(P0.05),对C2C12肌细胞的生肌分化有一定程度的促进作用。免疫印迹杂交结果显示MYOG和MYOD的蛋白质表达水平也发生了与基因表达相一致的变化。以上结果表明,CLA对动物骨骼肌细胞的正常生肌分化和生脂转分化都具有重要的调节作用。     

赤霉酸及矮壮素对苦瓜性别表现的影响

以兰山大白、株洲长白和英引３个苦瓜品种为材料进行外源激素处理对苦瓜植株性别表现影响的试验．结果表明，苗期进行ＧＡ３２５～１００ｍｇ／Ｌ处理有明显的促雌效果；ＣＣＣ５０～２００ｍｇ／Ｌ处理则有促雄效果，但促雄程度因品种而异，ＣＣＣ５００ｍｇ／Ｌ处理对兰山大白有适度的促雌作用．外源激素处理对苦瓜植株性别分化的影响可持续８０ｄ以上．苦瓜第一雌花形成与茎尖ＧＡ３含量达到峰值有关，２５ｍｇ／Ｌ的ＧＡ３处理使茎尖ＧＡ３含量的峰值提前出现而使第一雌花形成提早，而１００ｍｇ／ＬＣＣＣ处理使茎尖ＧＡ３含量先下降再逐渐回升而使植株第一雌花形成滞后