山羊TNNT3基因可变剪切及其对骨骼肌细胞分化的作用

【目的】快速骨骼肌肌钙蛋白T（fast skeletal troponin T3,TNNT3）作为肌钙蛋白(troponin, Tn)家族成员,调节横纹肌收缩、参与骨骼肌的生长发育并影响家畜肉质性状。通过获得山羊TNNT3基因的可变剪切体,分析山羊TNNT3基因可变剪切的表达模式及其在肌细胞分化中的作用,深入解析TNNT3基因在山羊骨骼肌生长发育过程中的作用机制。【方法】基于NCBI已公布山羊TNNT3基因（NM_001314210.1）和牛TNNT3基因（XM_010821200）mRNA序列,使用软件Primer Premier 6.0设计引物,以简州大耳羊胚胎期和出生后7个阶段骨骼肌为试验材料,克隆测序获得山羊TNNT3基因的CDS区可变剪切体,利用软件ORF Finder、EditSeq、DNAMAN、ClustalW和MEGA_X_10.1.8等对序列进行生物信息学分析;进一步设计实时荧光定量（real-time PCR,RT-qPCR）及半定量引物,研究TNNT3基因剪切体在7个不同组织（背最长肌（longissimus dorsi muscle,LD）、半膜肌（semimembranosus muscle,SM）、心、肝、脾、肺、肾）和7个发育阶段（胚胎期E75、E90、E105和出生后B3、B45、B150、B300）肌肉组织（背最长肌和半膜肌）中表达模式;此外,对转录本TNNT3_3进行体外编码能力检测确定其具有编码蛋白的能力,并在山羊骨骼肌卫星细胞（skeletal muscle satellite cells,MuSCs）中过表达,观察细胞形态变化以及检测标志基因的表达变化,研究其对山羊MuSCs分化的作用。【结果】①TNNT3（NM_001314210.1）CDS区全序列主要含有18个外显子,其中外显子16/17相互排斥,转录后单一表达。克隆发现山羊TNNT3基因 5个新转录本（TNNT3_1—5）,其外显子数分别是15、15、20、16、14。②生物信息学分析结果显示山羊TNNT3基因核苷酸序列和氨基酸序列与绵羊、牛、猪等哺乳动物具有很高的一致性,而与鱼类和爬行类动物的一致性较低,说明TNNT3基因序列在哺乳动物高度保守。③TNNT3 mRNA在背最长肌、半膜肌、心、肝、脾、肺、肾7个组织中都有表达,其中在骨骼肌中高度富集(P < 0.01),心脏及肺次之,其余组织中较低;TNNT3 mRNA在背最长肌和半膜肌中的表达始终处于一个动态变化中,胚胎期TNNT3在半膜肌的表达量高于背最长肌（P<0.05）;出生后则背最长肌中高于半膜肌（P<0.05）。④山羊TNNT3基因转录本TNNT3_3重复出现保守的外显子9—11（138bp）,体外翻译实验显示其可编码蛋白且蛋白大小与预期基本相符（37 kD）;相较于对照组,在山羊MuSCs中过表达该转录本使肌分化标志基因Myomaker、MyoG和MyH4 mRNA极显著升高(P < 0.01)。【结论】获得了山羊TNNT3基因具有完整CDS区5个新可变剪切体,TNNT3主要在肌肉组织（背最长肌和半膜肌）中高表达,在哺乳动物中高度保守且促进成肌分化。初步表明TNNT3基因在动物肌肉生长发育中具有重要的生物学功能。     

Habitat associations of saplings and adults in an old-growth temperate forest in the Changbai mountains,northeastern China

Species-habitat association analysis is useful to detect spatial arrangement of individual plants, to discover rules about the distribution of species and to generate hypotheses about the possible underlying process controlling observed structures. Quantifying methods were used to classify habitats in terms of topographical variables in a mixed temperate broad-leaved Korean pine forest of the Changbai mountains in northeastern China. All of the 625 20 m × 20 m quadrats of the plot could be unambiguously assigned to one of three habitat categories (low-plateau, high-plateau and slope). Torus-translation tests were used to estimate species-habitat associations. Many species are clearly distributed in a biased fashion with respect to habitats. Fifteen (55.6%) out of 27 species showed strong positive or negative association with specific habitats. We compared species-habitat associations at the sapling and adult stages. Adjusted density values indicated few species exhibit extremely strong habitat associations. Only 9 out of 26 species had adjusted densities > 3 in the habitat for which they had strong positive affinity. Few species show the same associations at the small tree and large tree stages. Only 3 out of 22 occurring associations with a specific habitat appeared to have a consistent habitat association at the two stages. These results suggest that species-habitat associations exist in the 25-ha plot of the temperate forest of the Chang- bai mountains. Owing to limitations in our statistical methodology, we partly underestimated associations by ignoring rare species. Regeneration niches can contribute to co-existence, but regeneration niches due to habitat associations play a limited role in species co-existence, since most species show a similar trend in habitat associations at the sapling and adult stages. We should pay more attention to shifts in habitat associations, i.e. niche shifts at different stages of existence.     

中金系列杨树的组培技术研究

中金系列杨树是杨柳科杨属白杨派植物,选择其腋芽（或顶芽）、嫩茎段、嫩叶片等为外植体,进行愈伤组织诱导试验、不定芽分化试验、生根试验,筛选出了适合植株分化、生根的培养基成分;对生根试管苗进行移栽基质配比试验,确定组培苗最佳生长环境。总结出适合中金系列杨树组培的技术流程,保证了杨树的遗传稳定性,降低了培养成本,加快了培养速度。     

马铃薯不同品种及高经济高营养“黑美人”脱毒培养技术的研究

解决马铃薯种薯退化问题一般采用茎尖组织脱毒培养(又叫分生组织脱毒培养).本实验又通过对马铃薯不同品种块茎具芽眼部分进行脱毒培养,培育出无病毒种苗,通过测其株高、茎粗、统计子叶数目、观察子叶颜色对比“黑美人”、新大品、土豆-10、陇薯3号生长势强弱.对以上两种脱毒培养制定消毒时间梯度,总结最佳消毒时间,分析污染原因并提出...     

Seasonal patterns of photochemical capacity and spring phenology reveal genetic differentiation among native Scots pine (Pinus sylvestris L.) populations in Scotland

Environment-driven genetic differentiation among populations is a common feature among forest trees, and an understanding of how populations have adapted to their home site conditions is essential for management and conservation practices. In Scotland, 84 native Scots pine (Pinus sylvestris L.) woodlands are recognised by the Forestry Commission and they occupy highly diverse environments from the maritime west coast to continental sites in eastern Scotland. However, it is not known whether adaptations to local environments along sharp temperature and rainfall gradients have occurred in different populations and as a result, the seed transfer guidelines of the species are based only on data from isozymes and monoterpenes. In this study of an outdoor common-garden trial, we used chlorophyll fluorescence to examine whether seedlings from 32 open-pollinated families and eight populations from sites experiencing contrasting annual temperature regimes differed in their response to variation in natural outdoor temperatures between September 2009 and May 2010. In addition, growth initiation in spring was recorded. Photochemical capacity at photosystem II F_v/F_m showed a distinct seasonal trend and remained at relatively high levels (∼0.7) until November. Following a period of over 2 weeks with temperatures below or close to 0 °C, F_v/F_m started decreasing towards its minimum values recorded in early March when population means varied between 0.35 and 0.45. By early May and along with rising temperatures, photochemical capacity had recovered to the same level as observed in early November. Populations were found to respond differently to the cold period starting in December. The largest drop in photochemical capacity was observed in seedlings from a low-altitude population located in the maritime western Scotland, while in seedlings from higher-altitude locations in the cooler eastern Scotland, the response was smaller. In March, the recovery of photochemical capacity was slowest in seedlings from the mildest and coolest sites. Evidence of adaptive genetic differentiation was also found in spring phenology. Initiation of shoot elongation and needle flush were earlier in families from higher altitudes (cooler areas), but population differences were not significant at the α = 0.05 level. These results suggest that adaptation to the spatially heterogeneous environment in Scotland has taken place in Scots pine and that in order to minimise the risk of planting maladapted seed stock, the patterns of environmental and adaptive genetic variation should be taken into account in the management of genetic resources in this species.     

铁皮石斛茎段丛生芽诱导研究

通过对铁皮石斛组培过程的外植体消毒方法,以及不同培养基、培养基添加物的筛选试验,结果表明:多种消毒剂配合使用并进行二次消毒的效果最好;基础培养基以1/2MS较好;1/2 MS+5.0mg/L NAA+2.0mg/L 6-BA是最适合丛芽分化的诱导培养基;在培养基中添加香蕉10%+洋芋10%生根多,瓶苗长势好。     

橡胶树不同类型芽片的嫁接繁殖试验

通过以橡胶树品种云研77-4、云研77-2的不同类型的褐色芽片、绿色芽片芽接试验表明:绿色芽片的芽接成活率和抽芽率明显高于褐色芽片;成活率高达96.3%～98.3%,抽芽率高达98.0%～98.3%,苗圃出苗率为92.4%～96.3%,而褐色芽片的成活率、抽芽率和苗圃出苗率仅为39.6%～42.6%、32.0%～37.0%和12.7%～15.8%。     

枣不定芽再生体系的研究

以枣优良品种骏枣为试材,研究了多种因素对其离体叶片及大田幼嫩茎段再生效率的影响.结果表明,幼嫩茎段再生较容易.叶片再生与生长调节物质种类和浓度及叶片成熟度有关.枣叶片不定芽再生的最佳培养基为1/2MS(大量元素减半)+TDZ0.5mg/L+IBA0.2m/L.不定芽再生率以上部叶片最高.     

促芽肥与杂交中稻再生力关系及其作用机理

１９９１－１９９３年以２个杂交中稻品种为材料，研究了促芽肥与杂交中稻再生力关系及其作用机理。结果表明，促芽肥不能提高到２、倒３节位活芽率，可提高倒４、５位活芽率。促芽肥对再生力的效果与母茎 鞘干物质重有关，当母茎鞘干物质过大或过小时，促芽肥的作用极小。