基于微卫星DNA的神农架川金丝猴遗传结构研究

川金丝猴(Rhinopithecus roxellana)是我国特有珍稀濒危物种,了解其种群遗传结构和关键影响因素,对该物种的保护具有重要意义。以我国分布最东端的湖北神农架川金丝猴种群为研究对象,基于非损伤性DNA技术和微卫星DNA遗传标记等分子生物学方法及景观遗传参数,探讨了神农架川金丝猴的遗传多样性和遗传结构,旨在为川金丝猴的研究及川金丝猴种群的可持续发展提供理论基础。利用12个多态性微卫星位点,在455份川金丝猴粪便样品中,共检测到62个微卫星等位基因;共鉴定出316个不同川金丝猴个体;种群的平均期望杂合度、平均观察杂合度和多态性信息含量分别为0.626、0.559和0.650;群体间的Nei's遗传距离为0.046—0.139,分化系数为0.015—0.046。结果表明与其他地区川金丝猴种群相比,神农架川金丝猴种群具有较低的遗传多样性水平,种群内部存在遗传分化趋势;结合景观参数分析表明地理距离不是影响神农架川金丝猴群体间遗传距离的主要因素,而生境中的灌丛和草地以及人类活动干扰可能是影响川金丝猴遗传交流的主要因素。     

人为干扰对神农架川金丝猴连通性及遗传多样性的影响

川金丝猴湖北种群是川金丝猴分布的最东缘,现主要分布于神农架国家公园和巴东自然保护区,其作为单独的管理单元,相对较低的遗传多样性、孤立的遗传状态和较小的种群数量,使得该种群面对环境变化脆弱性高。以神农架为研究区域,首先基于物种分布模型获得川金丝猴移动的阻力图层,利用电路理论分析适宜生境连接度;其次,设置道路和旅游两个人为干扰情景,运用最小费用距离模型与电路理论模型,分析道路和旅游活动对川金丝猴移动路径以及对不同亚群内和亚群间连通性的影响;最后,利用CDPOP软件模拟道路对川金丝猴遗传多样性的影响。结果表明:(1)神农架川金丝猴适宜生境分布较集中,生境斑块间存在多个潜在扩散区域;(2)道路改变了川金丝猴往来于南北生境间的最优路径,旅游干扰对川金丝猴移动路径无明显影响,其干扰主要在于川金丝猴对其具有回避性,回避距离为100 m;(3)道路显著降低金猴岭亚群内的连通性,并显著增加了所有亚群间的移动阻力,旅游活动对亚群内和亚群间的连通性无显著影响;(4)川金丝猴种群在150年内的遗传多样性呈下降趋势,道路的影响将加剧遗传多样性下降的程度。基于以上结果本文提出了针对性的保护建议,对于该物种的保护和其他濒危物种的类似研究具有较强的参考价值和借鉴意义。     

利用非损伤性方法评估神农架保护区川金丝猴种群遗传多样性

湖北神农架国家级自然保护区是中国川金丝猴(Rhinopithecus roxellana)的重要分布区之一,为了更好地了解神农架川金丝猴种群遗传结构,并制定有效的保护对策,采集了该保护区2个川金丝猴种群126份样品,其中分布在大龙潭的人工补食种群粪便样品60份,分布在千家坪的一个小种群粪便样品63份、肌肉样品3份。通过应用线粒体DNA(mtDNA)引物和筛选出的9对近缘微卫星位点成功地进行了川金丝猴种群的遗传多样性和遗传结构分析,同时使用Y染色体相关标记方法进行性别鉴定。结果显示:在24份川金丝猴mtDNA D-loop区401bp序列中,共检测出27个变异位点,定义了20个单倍型,单倍型多样性(h)为0.9820,核苷酸多样性(π)为0.0129;其中有9对微卫星位点能够在117份粪便样品DNA中稳定扩增,2对位点可能偏离Hardy-Weinberg平衡,平均有效等位基因数(Ne)为3.85,遗传杂合度(Ho)为0.7450;多态信息含量(PIC)为0.5950。使用Y染色体标记方法,在58个特有的基因型中检测到18个雄性和40个雌性川金丝猴。微卫星标记和mtDNA D-loop区基因序列均表明神农架川金丝猴存在较丰富的遗传多样性,同时2个取样点川金丝猴种群间出现了明显的遗传分化。     

神农架川金丝猴人工补食群的遗传多样性和亲缘关系

以湖北神农架国家级自然保护区的川金丝猴人工补食群体为研究对象,利用微卫星遗传标记对该群体的遗传多样性和亲缘关系进行研究,并对其种群的遗传多样性发展趋势进行了预测。结果显示,12个微卫星位点在该补食群体中共检测到50个等位基因,平均观察杂合度和平均期望杂合度分别为0.668和0.630,多态信息含量为0.442-0.696,平均0.567。亲权鉴定中共鉴定出11个父-母-子单元,各单元未有近亲繁殖的状况。亲缘系数结果显示具有亲缘关系的个体对达21.64%,群体平均亲缘系数为0.1108,现6个家庭单元中有两对候选亲本具有亲缘关系,表明该群体存在较高的近交风险。遗传多样性变化趋势的模拟结果表明,种群的数量对遗传多样性的影响比较大,群体中雄雌比例也对遗传多样性的损失有影响;为避免近亲交配,保持群体的遗传多样性水平,建议进一步做好人工补食群金丝猴的个体识别,完善家庭系谱,明确繁殖个体的遗传背景;引入一批有效的建群者来增加种群的数量及优化雄雌的比例。     

秦岭大坪峪川金丝猴(Rhinopithecus roxellana)种群生存力分析

小种群的保护已成为保护生物学中一个重要的问题,鉴于小种群极易灭绝的特性,寻找制约小种群存活的关键因子,形成快速拯救的措施,是目前对于小种群进行保护十分迫切的任务。根据秦岭大坪峪地区2个川金丝猴(Rhinopithecus roxellana)种群连续5a的观察数据,参照群体大小、配偶体制、迁移扩散、环境容纳量、死亡率、灾害的发生频率等种群参数,并结合川金丝猴其他种群的研究结果,利用旋涡模型(Vortex 10.0),对猴群未来100a内的数量动态进行了模拟分析。结果表明,在没有个体迁移的情况下,大坪峪种群种群100a间灭绝概率为95.8%和93%;当存在个体迁移和扩散的情况,灭绝概率为2%和0.4%。因此种群间个体迁移是秦岭大坪峪川金丝猴种群数量动态的限制因子。鉴于此,增加川金丝猴大坪峪种群间的个体交流,能够保证本群川金丝猴长期生存所需的遗传多样性。同时,增加秦岭川金丝猴各种群间的个体交流,建立群间的生境走廊对这一世界性濒危物种的长期存活具有重要的现实意义。     

川金丝猴遗传多样性研究进展

川金丝猴目前处于生境破碎、种群隔离的濒危境地,对其进行遗传多样性研究是有效保护这一物种的必要前提.结合文献,比较了川金丝猴遗传研究中常用样品采集方法、标记方法等,总结了其遗传多样性的研究现状,并建议通过建立遗传谱系来避免或减少近交,建立生态走廊促进群间基因交流,以提高其遗传变异能力.     

气候变化背景下湖北川金丝猴种群生存力分析

气候变化预计将改变野生动物的潜在适宜生境面积、分布与格局,从而加剧濒危物种的灭绝风险。湖北川金丝猴种群(Rhinopithecus roxellana)相对较低的遗传多样性、孤立的遗传状态和较小的种群数量,使得该种群面对环境变化具有更高的脆弱性。研究该种群在气候变化下的生境变化及种群动态,可为今后制定气候变化下的适应性对策提供参考。本文基于湖北川金丝猴分布点、生物气候变量及环境变量,建立川金丝猴物种分布模型,评估气候变化对川金丝猴未来(2050s)生境及环境容纳量的影响;结合川金丝猴种群参数,利用漩涡模型预测100年内,川金丝猴受到气候变化及极端天气灾害影响下的种群生存力状况。结果显示,在气候变化下,2050s川金丝猴适宜生境相比当前减少64.45%,大龙潭亚群、金猴岭亚群和千家坪亚群的环境容纳量相比当前分别下降93.48%、72.86%和13.96%;当前情景下,湖北川金丝猴在100年内种群数量能够保持逐渐增长,说明该种群是一个具有一定繁殖力的种群;极端天气灾害和气候变化均能造成湖北川金丝猴种群数量不同程度下降,二者共同作用时影响最大;不同亚群对极端天气灾害和气候变化的敏感性不同。因此,保护适宜生境、保证生境间连通性及提供极端天气灾害的干预保护是湖北川金丝猴种群保护的重要途径。     

神农架川金丝猴投食群的社会行为谱

为探讨人工投食条件下川金丝猴的社会行为类型,2007年1月-2008年10月,采用行为取样法和全事件记录法对湖北神农架自然保护区投食猴群的社会行为进行观察。确定投食群有45种典型社会行为,其中打架、抢食、抱腰、吻背、爬伏、邀配与以往对笼养和野生川金丝猴的研究有差别。     

人天然免疫基因TRIM5α的序列及进化分析

目的：克隆人TRIM5α基因,对其编码序列进行分析,并进行分子进化研究。方法：以HeLa细胞cDNA为模板,用RT-PCR方法克隆人TRIM5α基因,与人基因组序列对比分析其结构;用BioEdit、Genedoc和MEGA4.1软件进行蛋白序列联配和进化分析。结果：扩增了长1482bp的人TRIM5α基因片段,序列分析表明其覆盖了完整编码框,编码由493个氨基酸残基组成的人TRIM5α蛋白;人TRIM5α蛋白与黑猩猩、大猩猩、猩猩、猕猴TRIM5α蛋白的相似度分别为98.2%、96.8%、93.7%和87.6%。结论：克隆了人TRIM5α基因,为进一步研究该基因的功能奠定了基础。     

川金丝猴圈养种群现状分析

川金丝猴(Rhinopithecus roxellartae)圈养种群大多饲养在中国的动物园中。根据《川金丝猴国际谱系簿2002》记录,到2002年12月31日种群存活数量185只,包括野外捕获个体和圈养出生个体。近年圈养出生数量一直保持增加,到2000年超过野外捕获个体数量,现在种群的增加更多地依靠圈养出生个体数量的增加。近12年中种群繁殖率不断提高。每年新生幼仔中野外捕获个体组产仔比率逐年减少,圈养出生个体组产仔比率逐年增大,但到2002年前者仍高于后者40％。两组动物的繁殖率都有上升趋势,野外捕获个体组的繁殖率大多高于圈养出生个体组的繁殖率,二者有明显差异。种群遗传状况是基因多样性保存量较高,各小种群的基因多样性却处于较整体低的水平。提高子代的繁殖率,增加各机构之间的合作繁殖以提高小种群的基因多样性保持量,这两者对于种群的健康发展是非常重要的。     

Role for dopamine in malonate-induced damage in vivo in striatum and in vitro in mesencephalic cultures

Defects in mitochondrial energy metabolism have been implicated in the pathology of several neurodegenerative disorders. In addition, the reactive metabolites generated from the metabolism and oxidation of the neurotransmitter dopamine (DA) are thought to contribute to the damage to neurons of the basal ganglia. We have previously demonstrated that infusions of the metabolic inhibitor malonate into the striata of mice or rats produce degeneration of DA nerve terminals. In the present studies, we demonstrate that an intrastriatal infusion of malonate induces a substantial increase in DA efflux in awake, behaving mice as measured by in vivo microdialysis. Furthermore, pretreatment of mice with tetrabenazine (TBZ) or the TBZ analogue Ro 4-1284 (Ro-4), compounds that reversibly inhibit the vesicular storage of DA, attenuates the malonate-induced DA efflux as well as the damage to DA nerve terminals. Consistent with these findings, the damage to both DA and GABA neurons in mesencephalic cultures by malonate exposure was attenuated by pretreatment with TBZ or Ro-4. Treatment with these compounds did not affect the formation of free radicals or the inhibition of oxidative phosphorylation resulting from malonate exposure alone. Our data suggest that DA plays an important role in the neurotoxicity produced by malonate. These findings provide direct evidence that inhibition of succinate dehydrogenase causes an increase in extracellular DA levels and indicate that bioenergetic defects may contribute to the pathogenesis of chronic neurodegenerative diseases through a mechanism involving DA.     

2018年中国植物科学若干领域重要研究进展

2018年中国植物科学继续呈现快速发展的态势, 我国科学家在国际植物科学主流学术刊物发表论文数量大幅增加, 取得了多项具有重要影响的成果。调控植物生长-代谢平衡实现可持续农业发展入选2018年度中国科学十大进展; 中国被子植物区系进化历史研究入选2018年度中国生命科学十大进展。以水稻为代表的农作物和果蔬等经济作物研究在国际上已呈现出明显的优势, 若干领域已从“追赶”状态跨越到“领跑”地位。该文对2018年中国科学家在植物科学若干领域取得的重要研究成果进行了概括性评述, 旨在全面追踪和报道当前中国植物科学领域的发展前沿和热点, 展示中国科学家所取得的杰出成就。     

2017年中国植物科学若干领域重要研究进展

2017年中国植物科学继续保持高速发展态势, 重大成果频出, 具体表现在中国植物学家在国际顶级学术期刊发表的文章数量平稳上升。中国植物科学领域的研究工作者成果精彩纷呈, 如新型广谱抗病机制的发现、水稻广谱抗病遗传基础及机制和疫霉菌诱发病害成灾机制研究等。2017年中国生命科学领域十大进展评选中, 有两项植物科学领域的研究成果入选。水稻生物学、进化与基因组学和激素生物学等领域学科发展突出。另外, 值得一提的是, 长期从事高等植物与代谢途径调控分子网络研究和水稻品种设计育种的李家洋院士的研究成果“水稻高产优质性状形成的分子机理及品种设计”荣获2017年国家自然科学一等奖。这一具有重大国际影响的开创性贡献标志着中国植物科学在该领域的国际科学前沿居于引领和卓越地位。该文对2017年中国本土科学家在植物科学若干领域取得的重要研究成果进行了系统梳理, 旨在全面追踪和报道当前中国植物科学领域发展的最新前沿动态, 与广大读者共同分享我国科学家所取得的辉煌成就。     

SSTR5 ablation in islet results in alterations in glucose homeostasis in mice

Somatostatin (SST) peptide is a potent inhibitor of insulin secretion and its effect is mediated via somatostatin receptor 5 (SSTR5) in the endocrine pancreas. To investigate the consequences of gene ablation of SSTR5 in the mouse pancreas, we have generated a mouse model in which the SSTR5 gene was specifically knocked down in the pancreatic beta cells (betaSSTR5Kd) using the Cre-lox system. Immunohistochemistry analysis showed that SSTR5 gene expression was absent in beta cells at three months of age. At the time of gene ablation, betaSSTR5Kd mice demonstrated glucose intolerance with lack of insulin response and significantly reduced serum insulin levels. Insulin tolerance test demonstrated a significant increase of insulin clearance in vivo at the same age. In vitro studies demonstrated an absence of response to SST-28 stimulation in the betaSSTR5Kd mouse islet, which was associated with a significantly reduced SST expression level in betaSSTR5Kd mice pancreata. In addition, betaSSTR5Kd mice had significantly reduced serum glucose levels and increased serum insulin levels at 12 months of age. Glucose tolerance test at an older age also indicated a persistently higher insulin level in betaSSTR5Kd mice. Further studies of betaSSTR5Kd mice had revealed elevated serum C-peptide levels at both 3 and 12 months of age, suggesting that these mice are capable of producing and releasing insulin to the periphery. These results support the hypothesis that SSTR5 plays a pivotal role in the regulation of insulin secretion in the mouse pancreas.     

2018年中国植物科学若干领域重要研究进展

2018年中国植物科学继续呈现快速发展的态势, 我国科学家在国际植物科学主流学术刊物发表论文数量大幅增加, 取得了多项具有重要影响的成果。调控植物生长-代谢平衡实现可持续农业发展入选2018年度中国科学十大进展; 中国被子植物区系进化历史研究入选2018年度中国生命科学十大进展。以水稻为代表的农作物和果蔬等经济作物研究在国际上已呈现出明显的优势, 若干领域已从“追赶”状态跨越到“领跑”地位。该文对2018年中国科学家在植物科学若干领域取得的重要研究成果进行了概括性评述, 旨在全面追踪和报道当前中国植物科学领域的发展前沿和热点, 展示中国科学家所取得的杰出成就。     

我国葫芦科植物离体培养研究进展

葫芦科植物包括多种瓜类蔬菜,对其进行离体培养研究具有重要的理论和实践意义.综述了国内在葫芦科植物器官培养、体细胞胚胎发生、花药培养、原生质体培养和体细胞杂交及离体遗传转化等方面取得的研究进展,并对葫芦科植物离体培养、遗传转化与育种的前景作了展望.     

我国葫芦科植物离体培养研究进展

葫芦科植物包括多种瓜类蔬菜,对其进行离体培养研究具有重要的理论和实践意义。综述了国内在葫芦科植物器官培养、体细胞胚胎发生、花药培养、原生质体培养和体细胞杂交及离体遗传转化等方面取得的研究进展,并对葫芦科植物离体培养、遗传转化与育种的前景作了展望。     

Changes in aquatic vegetation in Rhine floodplain streams in Alsace in relation to disturbance

Abstract. Changes are described in aquatic vegetation in oligotrophic, groundwater-fed Rhine floodplain streams in Alsace (eastern France), resulting from disturbance. Disturbance factors include changes in nutrients, either permanent ones - effluent from a waste water treatment plant or trout hatcheries - or periodic ones: flooding. Regular inputs of high levels of phosphate and ammonia modified the macrophyte vegetation in these streams. The floristic composition, which was characteristic of oligotrophic waters upstream of the eutrophicated sector, changed to that of a eutrophic situation as originally found downstream. Periodic disturbance by floods which normally occur once a year, irregularly eutrophicates the small streams, causing the development of a mixture of eutrophic and oligotrophic species. Six macrophyte communities are distinguished, indicating different trophic levels. The aquatic vegetation is adapted to the variations of phosphate and ammonia levels. Hence, aquatic macrophytes can be used as bio-indicators of fluctuations in water nutrient levels in relation to the type of disturbance.     

Apoptosis in experimental myocardial infarction in situ and in the perfused heart in vitro

We report the appearance of apoptotic cells in experimental myocardial infarction (rabbit heart) in in situ and in vitro preparations. Apoptosis was recognized by intravital staining with Hoechst 33342 (Ho342), by nick-end labeling (TUNEL) and by DNA laddering. A steady rise in the relative number of apoptotic cardiomyocytes (apoptotic index) was noted in in situ preparations. Apoptosis was first noted 6 h after the onset of ischemia with its highest value occurring after 72 h. Apoptotic nuclei were absent in remote areas of the left and right ventricles. Apoptotic nuclei within the infarcted area showed diminished intensity of Ho342 fluorescence. Three days after ischemia, a border zone adjacent to the infarcted area consisting of apoptotic macrophages was recognized. A novel finding was the appearance of apoptotic cardiomyocytes in the isolated perfused ischemic heart. Occurring as early as 50 min after the onset of ischemia, a high apoptotic index was present adjacent to the ligature placed around the coronary artery. This observation provides the opportunity to selectively examine factors leading to apoptosis in the ischemic heart under controlled experimental conditions.     

Morphine-potentiated platelet aggregation in in vitro and platelet plug formation in in vivo experiments

The detailed mechanisms underlying morphine-signaling pathways in platelets remain obscure. Therefore, we systematically examined the influence of morphine on washed human platelets. In this study, washed human platelet suspensions were used for in vitro studies. Furthermore, platelet thrombus formation induced by irradiation of mesenteric venules with filtered light in mice pretreated with fluorescein sodium was used for an in vivo thrombotic study. Morphine concentration dependently (0.6, 1, and 5 microM) potentiated platelet aggregation and the ATP release reaction stimulated by agonists (i.e., collagen and U46619) in washed human platelets. Yohimbine (0.1 microM), a specific alpha(2)-adrenoceptor antagonist, markedly abolished the potentiation of morphine in platelet aggregation stimulated by agonists. Morphine also potentiated phosphoinositide breakdown and intracellular Ca(2+) mobilization in human platelets stimulated by collagen (1 microg/ml). Moreover, morphine (0.6-5 microM) markedly inhibited prostaglandin E(1) (10 microM)-induced cyclic AMP formation in human platelets, while yohimbine (0.1 microM) significantly reversed the inhibition of cyclic AMP by morphine (0.6 and 1 microM) in this study. The thrombin-evoked increase in pH(i) was markedly potentiated in the presence of morphine (1 and 5 microM). Morphine (2 and 5 mg/g) significantly shortened the time require to induce platelet plug formation in mesenteric venules. We concluded that morphine may exert its potentiation in platelet aggregation by binding to alpha(2)-adrenoceptors in human platelets, with a resulting inhibition of adenylate cyclase, thereby reducing intracellular cyclic AMP formation followed by increased activation of phospholipase C and the Na(+)/H(+) exchanger. This leads to increased intracellular Ca(2+) mobilization, and finally potentiation of platelet aggregation and of the ATP release reaction.