种牛精液冷冻和保存效果研究进展

随着养牛业的高速发展,牛精液液体保存技术的研究也在逐步深入。人工授精技术降低了养殖成本,加速了肉牛的繁殖改良,同时还促进了育种工作的进程,合理有效的使用人工授精技术能够提高牛的饲养效益。与此同时,优良的种牛精液品质也是改良的关键,有实验结果表明：遗传、气候环境、饲养管理等因素会影响牛的精液品质[1],同时强制运动也是决定种牛精液品质的关键环节[2]。合理的精液冷冻与保存不仅能发挥优良的精液品质,同时也为人工授精技术的发展提供保障。本文从牛精液新型冷冻保护剂和损伤修复、牛精液冷冻保存添加剂、牛冷冻精在人工受精中的应用、冷冻保存的发展前景等方面总结了我国牛精液冷冻保存技术的研究和发展,以期对今后种牛精液冷冻和保存技术发展有所帮助、提供参考。     

‘岩溪晚芦’椪柑果皮与果肉差减cDNA文库的构建及初步分析

 以‘岩溪晚芦’(Citrus reticulata Blanco)成熟果皮和果肉为材料,采用抑制性差减杂交技术,成功构建了果皮（tester）与果肉（driver）的差减cDNA文库。结果显示：cDNA克隆外源插入片段大小介于100～500 bp之间。成功对411个克隆进行了测序,测序结果经与NCBI基因库中序列进行比较,377个EST 找到了同源序列,它们分属于126个基因,涉及到与色素合成、能量代谢、初级代谢、次生代谢、抗逆防御、蛋白代谢、信号转导、香味形成、果皮软化（衰老）、精油合成、抗氧化代谢等代谢途径和生理生化过程。还有一些基因涉及到三价铁螯合物、反转录转座子gag蛋白、查尔酮合成、硼离子转运、儿茶酚氧位甲基转移酶等功能。功能已知且没有重复的EST共有63个,占总数的49.2%。有5个基因共62个克隆虽在Blast搜索时找到了较高的同源性序列,但功能目前尚未明确,有待进一步深入分析。另有34个基因未搜索到同源序列,可能为新发现的基因。     

基于LAN的农业科学研究院的科研仪器共享共用信息平台设计与实现

为提高科研院所大型仪器的利用率、降低科研成本,以浙江省农业科学院大型仪器信息化管理为研发对象,研发了大型仪器管理系统,重点论述了该系统的设计思想、系统结构及主要功能。     

A rapid multiplication system for ‘Candidatus Liberibacter asiaticus’ through regeneration of axillary buds in vitro

‘Candidatus Liberibacter asiaticus (CLas)’, which causes citrus Huanglongbing (HLB) disease, has not been successfully cultured in vitro to date. Here, a rapid multiplication system for CLas was established through in vitro regeneration of axillary buds from CLas-infected ‘Changyecheng’ sweet orange (Citrus sinensis Osbeck). Stem segments with a single axillary bud were cultured in vitro to allow CLas to multiply in the regenerating axillary buds. A high CLas titer was detected in the regenerated shoots on an optimized medium at 30 days after germination (DAG). This titer was 28.2-fold higher than in the midribs from CLas-infected trees growing in the greenhouse. To minimize contamination during in vitro regeneration, CLas-infected axillary buds were micrografted onto seedlings of ‘Changyecheng’ sweet orange and cultured in a liquid medium. In this culture, the titers of CLas in regenerated shoots rapidly increased from 7.5×10⁴ to 1.4×10⁸ cells μg^–1 of citrus DNA during the first 40 DAG. The percentages of shoots with >1×10⁸ CLas cells μg^–1 DNA were 30 and 40% at 30 and 40 DAG, respectively. Direct tissue blot immunoassay (DTBIA) indicated that the distribution of CLas was much more uniform in regenerated plantlets than in CLas-infected trees growing in the greenhouse. The disease symptoms in the plantlets were die-back, stunted growth, leaf necrosis/yellowing, and defoliation. The death rate of the plantlets was 82.0% at 60 DAG. Our results show that CLas can effectively multiply in citrus plantlests in vitro. This method will be useful for studying plant-HLB interactions and for rapid screening of therapeutic compounds against CLas in citrus.     

四倍体小麦矮秆基因的赤霉素敏感性及对农艺性状的影响

为明确四倍体小麦矮秆基因Rht14、Rht16和Rht18的赤霉素敏感性及其对小麦农艺性状的影响,促进这些矮秆基因的合理利用。选用分别含有Rht14、Rht16和Rht18的四倍体小麦近等基因系ANW16D(Rht14)、ANW16F(Rht16)、ANW16G(Rht18)及其高秆轮回亲本LD222以及六倍体小麦中国春(Chinese spring),测量其不同浓度GA3处理下小麦的株高,计算赤霉素敏感系数(GRI)并推断3种矮秆小麦的赤霉素反应类型。在成熟期对LD222近等基因系小麦的农艺性状如株高、穗长、主穗穗下第一茎节(P-1)节间长、节间表皮细胞、种子表皮细胞及种子体积等进行测量,分析Rht14、Rht16、Rht18这3个矮秆基因对小麦这些农艺性状的效应。结果表明,ANW16D、ANW16F和ANW16G这3个矮秆小麦株高恢复到正常LD222株高的最适GA3浓度为10-4mol/L;3个矮秆品种均为赤霉素敏感型且敏感性大小为中国春ANW16GANW16FANW16DLD222;Rht14、Rht16、Rht18均是通过降低主穗穗下第一茎节(P-1)节间长度来使小麦株高降低,降低效应为Rht18Rht16Rht14;它们降低小麦株高的根本原因均是缩短了小麦节间表皮细胞长度且缩短效应与降低株高效应一致;3个矮秆基因均在降低小麦株高的同时不影响种子的体积。     

Effect of EZH2 regulating Wnt/β-catenin signaling pathway on apoptosis of brain glioma cells

AIM: To investigate the effect of enhancer of zeste homolog 2 (EZH2) regulating Wnt/β-catenin signaling pathway on the apoptosis of brain glioma cell lines. METHODS: The expression level of EZH2 in glioma cell lines U87, H4 and U251 and normal human astrocytes (NHA) was detected by RT-qPCR and Western blot. The EZH2 siRNA and siRNA control were transfected into the H4 cells. The cell viability was measured by MTT assay. The apoptosis was analyzed by flow cytometry. Caspase-3 activity was detected by spectrophotometry. The expression levels of the key protein β-catenin of the Wnt/β-catenin signaling pathway and the downstream target molecule c-Myc were determined by Western blot. After the H4 cells transfected with EZH2 siRNA were treated with an activator of Wnt/β-catenin signaling pathway, the apoptosis rate was measured by flow cytometry, and the expression of β-catenin and c-Myc was determined by Western blot. RESULTS: The mRNA and protein expression levels of EZH2 in the glioma cell lines U87, H4 and U251 were significantly higher than those in NHA (P<0.05). The expression of EZH2 at mRNA and protein levels in the H4 cells was higher than that in U87 cells and U251 cells (P<0.05). EZH2 siRNA obviously inhibited the expression of EZH2 at mRNA and protein levels in the H4 cells. Knockdown of EZH2 expression decreased the viability of H4 cells, the apoptotic rate was significantly increased, and the activity of caspase-3 was significantly increased in the cells (P<0.05). Knockdown of EZH2 expression also inhibited the expression of β-catenin and c-Myc. The activator of Wnt/β-catenin signaling pathway reduced the apoptosis rate of H4 cells induced by down-regulation of EZH2, and reduced the activity of caspase-3 in the cells. CONCLUSION: EZH2 is over-expressed in glioma cells. Down-regulation of EZH2 expression induces apoptosis of glioma cells by inhibiting the activation of Wnt/β-catenin signaling pathway.     

生长调节物质对楸树花粉萌发及花粉管生长的影响

采用离体花粉液体培养法研究4种植物生长调节物质对不同来源楸树花粉萌发及花粉管生长的影响。结果表明：较低浓度的赤霉素促进花粉萌发及花粉管生长,高浓度则表现为明显的抑制作用,最适浓度为300mg/L;吲哚乙酸和2,4-D对花粉的作用效果与楸树的种源有关,总体上均呈低浓度促进,高浓度抑制的规律;花粉萌发率均在吲哚乙酸为15mg/L时达到最大值,只有2个种源与对照间差异极显著,花粉萌发率在2,4-D为200mg/L时均达到最大值,超过最适浓度后,吲哚乙酸和2,4-D均对花粉萌发及花粉管生长表现为抑制作用;各个浓度的萘乙酸均抑制花粉萌发及花粉管生长,且抑制程度随浓度增加而加强。     

准葛尔盆地南缘沙尘暴、大风气候特征分析

选取1953~2006年准葛尔盆地南缘石河子垦区3个地面观测站的资料,对比分析了沙尘暴、大风的时空分布特征,探讨沙尘暴和大风之间的关系及沙尘暴发生时的最多风向和最小风速指标。结果表明:准葛尔盆地南缘石河子垦区沙尘暴与大风的地理分布刚好相反,沙尘暴多的地方大风较少,北部莫索湾沙漠边缘一带为沙尘暴高发区;但有着基本一致的气候变化趋势。沙尘暴发生时的最多风向为西北风,引起沙尘暴的最小风速指标大于10m/s。     

miR-21:生成、表达调控及对其细胞凋亡的影响

凋亡是细胞的一种基本生物学现象,在机体生命过程中发挥着重要作用。miRNAs是在真核生物内发现的一类长度约22个核苷酸的内源性非编码单链小RNA,通过调控靶基因的表达参与调控细胞的凋亡、分化、增殖、侵袭和代谢。大量的研究表明,miR-21作为miRNAs重要成员之一,与细胞凋亡有重要联系。本文将从miR-21的生成、表达调控及对细胞凋亡的影响方面进行综述。     

深圳市道路绿化管养标准体系构建浅析

城市道路绿化在改善城市生态环境、提升市民生活质量方面有重要作用。在道路绿化中,管养工作是维持道路绿化活力的重要内容。通过考察研究深圳市以及其他省市的道路绿化管养标准体系,分析对比每个城市的道路绿化管养标准体系现状,指出了深圳市道路绿化管养标准体系具有建立较早、内容较为全面的优势,以及存在体系架构不健全、部分标准标龄过长、可量化执行标准较少等方面的不足。并从系统性、指导性、先进性角度提出优化建议,将深圳市道路绿化管养标准体系划分为基础标准和道路绿化管养通用标准两大部分,其中基础标准分为术语标准等4个大类,道路绿化管养通用标准划分为质量技术等14个类别。