Genetic consequence of restricted habitat and population decline in endangered Isoetes sinensis (Isoetaceae)

BACKGROUND AND AIMS: Isoetes sinensis (Isoeteaceae) is a critically endangered aquatic quillwort in eastern China. Rapid decline of extant population size and local population extinction have occurred in recent years and have raised great concerns among conservationists. METHODS: Amplified fragment length polymorphisms (AFLPs) were used to investigate the genetic variation and population structure of seven extant populations of the species. KEY RESULTS: Eight primer combinations produced a total of 343 unambiguous bands of which 210 (61.2 %) were polymorphic. Isoetes sinensis exhibited a high level of intra-population genetic diversity (H(E) = 0.118; hs = 0.147; I = 0.192; P = 35.2 %). The genetic variation within each of the populations was not positively correlated with their size, suggesting recent population decline, which is well in accordance with field data of demographic surveys. Moreover, a high degree of genetic differentiation (F(ST) = 0.535; G(ST) = 0.608; theta(B) = 0.607) was detected among populations and no correlation was found between geographical and genetic distance, suggesting that populations were in disequilibrium of migration-drift. Genetic drift played a more important role than gene flow in the current population genetic structure of I. sinensis because migration of I. sinensis is predominantly water-mediated and habitat range was highly influenced by environment changes. CONCLUSIONS: Genetic information obtained in the present study provides useful baseline data for formulating conservation strategies. Conservation management, including both reinforcement for in situ populations and ex situ conservation programmes should be carefully designed to avoid the potential risk of outbreeding depression by admixture of individuals from different regions. However, translocation within the same regional population should be considered as a measure of genetic enhancement to rehabilitate local populations. An ex situ conservation strategy for conserving all extant populations to maximize genomic representation of the species is also recommended.     

Purification and Partial Characterization of β-Glucosidase from Fresh Leaves of Tea Plants （Camellia sinensis （L.） O. Kuntze）

β-Glucosidases are important in the formation of floral tea aroma and the development of resistance to pathogens and herbivores in tea plants. A novel β-glucosidase was purified 117-fold to homogeneity,with a yield of 1.26%, from tea leaves by chilled acetone and ammonium sulfate precipitation, ion exchange chromatography (CM-Sephadex C-50) and fast protein liquid chromatography (FPLC; Superdex 75, Resource S). The enzyme was a monomeric protein with specific activity of 2.57 U/mg. The molecular mass of the enzyme was estimated to be about 41 kDa and 34 kDa by SDS-PAGE and FPLC gel filtration on Superdex 200, respectively. The enzyme showed optimum activity at 50℃ and was stable at temperatures lower than 40℃. It was active between pH 4.0 and pH 7.0, with an optimum activity at pH 5.5, and was fairly stable from pH 4.5 to pH 8.0. The enzyme showed maximum activity towards pNPG, low activity towards pNP-Galacto, and no activity towards pNP-Xylo.     

我国东南部地区夏季两种雀形目鸟类的代谢产热特征及其体温调节

采用封闭式流体压力呼吸计,分别在5-35℃和5-40℃的环境温度范围内测定了白头鹎(Pycnonotussinensis)和丝光椋鸟(Sturnussericeus)的代谢率、热传导和体温等指标,探讨其代谢产热特征。结果显示:在环境温度(Ta)为5-35℃时,白头鹎的体温基本维持恒定,平均温度为40.3±0.1℃,热中性区为26.6-32.8℃,基础代谢率为73.10±4.11mlO2/h,是体重预期值的79%;Ta在5-26℃范围内,代谢率(MR)与Ta呈负相关,回归方程为:MR[mlO2/h]=265.37-7.24Ta(℃);Ta在5-30℃范围内,热传导值最低且基本保持恒定,平均为0.24±0.01mlO2/g·h·℃,是体重预期值的126%。丝光椋鸟的热中性区为27.6-34.5℃,平均体温为40.5±0.1℃(5-40℃),基础代谢率为160.64±9.20mlO2/h,是体重预期值的90%;最低热传导为0.16±0.05mlO2/g·h·℃,是体重预期值的129%。在5-25℃范围内,MR与Ta的回归方程为:MR[mlO2/h]=377.96-7.88Ta(℃)。白头鹎和丝光椋鸟的基本生物学特征为:较高的体温,热传导和上临界温度,较宽的热中性区和较低的代谢率,符合南方小型鸟类的代谢特征.     

茶树鲜花饮料澄清技术研究

茶树鲜花与茶叶具有相似的营养成分,榨汁后经调配可开发成适口性好、风味独特的鲜花饮料。对茶树鲜花饮料的壳聚糖絮凝、酶法澄清、超滤澄清效果进行了比较,结果表明,采用10万分子截留量的超滤膜处理获得能够较好的澄清效果,且对茶多酚造成的操作损失较小。     

冬虫夏草及其相关类群的分子系统学分析

为了探明冬虫夏草及其相关类群的亲缘关系,以冬虫夏草、中国被毛孢及中华束丝孢(=冬虫夏草头孢=蝙蝠蛾多毛孢)共6株菌种作为内群和一株蛹草拟青霉作为外群进行了DNA随机多态型(RAPD)分析。此外,基于上述供试材料又在内群中增加了一株蝙蝠蛾拟青霉,并对内群和外群样品的nrDNA间隔区(ITS)碱基序列进行了测定;对于测定的8条序列连同来自GenBank中的4条相关序列进行了分子系统学分析。结果表明:中华束丝孢和中国被毛孢均系冬虫夏草菌的无性型。按照国际植物命名法规,中国被毛孢应为冬虫夏草菌无性型的正确名称。而蝙蝠蛾拟青霉为不同于冬虫夏草菌的另一种真菌;该名称由于不合格发表而不被国际植物命名法规所承认。     

蝎蛉科与蚊蝎蛉科昆虫触角感器超微形态比较

触角是昆虫重要的感觉器官,其感觉功能主要是通过其上的各类感器来实现的。本文以染翅蝎蛉Sinopanorpa tincta(Nav偄s)和中华蚊蝎蛉Bittacus sinensis Walker为代表,采用扫描电子显微镜观察了蝎蛉科Panorpidae和蚊蝎蛉科Bittacidae触角感器的类型及其超微结构,并简要讨论了各种感器的功能。结果显示:染翅蝎蛉触角上共发现7种感器,分别为短刺形感器、Bhm氏鬃毛、毛形感器、短毛形感器、锥形感器、刺形感器、和微毛形感器。中华蚊蝎蛉触角上也发现7种感器,分别为Bhm氏鬃毛、芽形感器、毛形感器、刺形感器、畸形毛感器、腔锥感器和钟形感器。蚊蝎蛉触角与蝎蛉的最大区别是着生有大量的腔锥感器,初步推测众多的腔锥感器可能与蚊蝎蛉对湿度的高度敏感有关。     

克隆植物中国沙棘生长对外源植物激素的响应

关于植物克隆生长调节问题,目前集中于外在机制的研究。为了探讨中国沙棘克隆生长调节的内在机制,采用3×3回归设计进行田间试验,期望了解不同生长性状对IAA和CTK用量及其配比的响应规律。结果表明:(1)生长性状对激素用量的响应规律呈典型的钟形曲面模式,即各生长指标均存在一个产量峰值,峰值以前生长指标随IAA、CTK用量的增大而提高,峰值以后生长指标随IAA、CTK用量的增大而下降。(2)不同生长指标对激素用量及其配比的响应规律具有一定差异,较高的IAA比例有利于促进树高生长,较高的CTK比例有利于促进地径和冠幅生长,而几乎相等的IAA和CTK用量有利于种群生物量积累。(3)在激素用量适宜的情况下,中国沙棘生长潜力得到充分发挥,形成高大的个体,较多的子株,有利于提高种群对生境资源的占据和利用,并提高排斥其他植物种类入侵的能力;当激素用量过高或过低时,中国沙棘以降低生长量为代价,形成矮小的个体,减少子株数量,有利于削弱个体之间的竞争。这一结果为了解中国沙棘克隆生长内在调节机制提供了线索。(4)根据不同生长指标的激素效应方程,求出了相应的IAA和CTK的最佳用量和最佳配比,以及合理施激素区域和最低成本线。(5)克隆子株数量增幅与地径和冠幅生长量增幅呈极显著正相关、与种群生物量增幅呈显著正相关,即适宜的IAA和CTK用量既可加速个体生长、也能促进克隆子株的产生。     

雄全异株植物瘿椒树（省沽油科）的传粉生物学

从开花动态、传粉昆虫、花的形态结构、繁育系统、花粉活力和柱头可授性等方面研究了我国特有珍稀植物瘿椒树（Tapiscia sinensis Oliv.）的传粉生物学特性。瘿椒树是典型的雄全异株植物,两性花中含有功能性花粉,且自交亲和,但雄花花粉活力和萌发力是两性花的10倍以上。雄株和两性植株具有相同开花物候期,花期均为5月下旬至6月上旬,单花期为4-5天,雄花和两性花的5枚花药开裂的不同步性明显延长了散粉时间。两性花雌蕊先熟,柱头可授性较长。具有适应风媒和虫媒传粉的花部特征。传粉昆虫主要为蜜蜂科（Apidae）和食蚜蝇科（Syrphidae）昆虫,访花高峰期为8：30-10：30。维持瘿椒树雄全异株的可能机制是：雄株总体上增加了异交花粉的数量和质量;两性花的雄蕊为该物种提供了繁殖保障,同时为传粉者提供了报酬。     

不同产地中华鳖的线粒体控制区序列分析及结构比较

采用PCR特异引物,扩增了两产地中华鳖（Pelodiscus sinensis）个体的mtDNA控制区（CR）及其邻近片段,测序获得了长度分别为1830bp和1630bp的序列。结合GenBank中已发表的韩国产中华鳖mtDNA的CR区序列,比较了3个产地中华鳖的CR区结构。分析显示：中华鳖不同产地mtDNA CR区DNA中的A＋T含量分别为60.5%、63.6%和64.8%,它们的5′、3′末端以及CSB1-CSB2之间均存在丰富的可变数目串联重复序列（variable numbers oftandem repeats,VNTR）。基于mtDNA CR区序列和结构分析,显示中华鳖不同产地的野生个体中存在丰富的遗传多样性。     

Characterization of electrical penetration graphs of the Asian citrus psyllid, Diaphorina citri, in sweet orange seedlings

Detailed information on probing behavior of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is critical for understanding the transmission process of phloem‐limited bacteria (Candidatus Liberibacter spp.) associated with citrus ‘huanglongbing’ by this vector. In this study, we investigated stylet penetration activities of D. citri on seedlings of Citrus sinensis (L.) Osbeck cv. Pêra (Rutaceae) by using the electrical penetration graph (EPG‐DC system) technique. EPG waveforms were described based on amplitude, frequency, voltage level, and electrical origin of the observed traces during stylet penetration into plant tissues. The main waveforms were correlated with histological observations of salivary sheath termini in plant tissues, to determine the putative location of stylet tips. The behavioral activities were also inferred based on waveform similarities in relation to other Sternorrhyncha, particularly aphids and whiteflies. In addition, we correlated the occurrence of specific waveforms with the acquisition of the phloem‐limited bacterium Ca. Liberibacter asiaticus by D. citri. The occurrence of a G‐like xylem sap ingestion waveform in starved and unstarved psyllids was also compared. By analyzing 8‐h EPGs of adult females, five waveforms were described: (C) salivary sheath secretion and other stylet pathway activities; (D) first contact with phloem (distinct from other waveforms reported for Sternorrhyncha); (E1) putative salivation in phloem sieve tubes; (E2) phloem sap ingestion; and (G) probably xylem sap ingestion. Diaphorina citri initiates a probe with stylet pathway through epidermis and parenchyma (C). Interestingly, no potential drops were observed during the stylet pathway phase, as are usually recorded in aphids and other Sternorrhyncha. Once in C, D. citri shows a higher propensity to return to non‐probing than to start a phloem or xylem phase. Several probes are usually observed before the phloem phase; waveform D is observed upon phloem contact, always immediately followed by E1. After E1, D. citri either returns to pathway activity (C) or starts phloem sap ingestion, which was the longest activity observed.