Structural and Functional Analyses of β-Glucosidase 3B from Thermotoga neapolitana: A Thermostable Three-Domain Representative of Glycoside Hydrolase 3

Based on sequence and phylogenetic analyses, glycoside hydrolase (GH) family 3 can be divided into several clusters that differ in the length of their primary sequences. However, structural data on representatives of GH3 are still scarce, since only three of their structures are known and only one of them has been thoroughly characterized—that of an exohydrolase from barley. To allow a deeper structural understanding of the GH3 family, we have determined the crystal structure of the thermostable β-glucosidase from Thermotoga neapolitana, which has potentially important applications in environmentally friendly industrial biosynthesis at a resolution of 2.05 Å. Selected active-site mutants have been characterized kinetically, and the structure of the mutant D242A is presented at 2.1 Å resolution. Bgl3B from Th. neapolitana is the first example of a GH3 glucosidase with a three-domain structure. It is composed of an (α/β)₈ domain similar to a triose phosphate isomerase barrel, a five-stranded α/β sandwich domain (both of which are important for active-site organization), and a C-terminal fibronectin type III domain of unknown function. Remarkably, the direction of the second β-strand of the triose phosphate isomerase barrel domain is reversed, which has implications for the active-site shape. The active site, at the interface of domains 1 and 2, is much more open to solvent than the corresponding site in the structurally homologous enzyme from barley, and only the − 1 site is well defined. The structures, in combination with kinetic studies of active-site variants, allow the identification of essential catalytic residues (the nucleophile D242 and the acid/base E458), as well as other residues at the − 1 subsite, including D58 and W243, which, by mutagenesis, are shown to be important for substrate accommodation/interaction. The position of the fibronectin type III domain excludes a direct participation of this domain in the recognition of small substrates, although it may be involved in the anchoring of the enzyme on large polymeric substrates and in thermostability.     

Altitudinal variations in seedling and sapling density and age structure of timberline tree species in the Sergyemla Mountains, southeast Tibet

The growth situation of seedlings of treeline species can be used as an indicator for the upper or lower shift of treeline under global climate change, since any advance of a treeline would require seedling survival above the current treeline elevation. We investigated seedling and sapling density of two dominant conifers Abies georgei var. smithii and Sabina saltuaria at lower, middle and upper altitude of two contrasting slopes in Sergyemla Mountain, southeast Tibet. Seedling and sapling age structure of A. georgei var. smithii was analyzed further. Seedling density of both species, as well as sapling density of S. saltuaria, showed a declining trend as elevation increased. Sapling density of A. georgei var. smithii was somewhat higher at middle altitude than other positions. The frequency of old-aged A. georgei var. smithii seedlings/saplings (age >10 year) was the lowest at the highest altitude and tended to increase with the declining elevation. The results indicate that the higher density of seedling and sapling in high-altitude might be attributed to better soil moisture availability and solar insolation than in the mid- and low-altitudes during the growth season. Yet confined by other environmental factors, A. georgei var. smithii seedlings in high-altitude suffer from lower survival rate and lower possibility to grow tall compared with those in mid- and low-altitudes.     

野生荆条籽中挥发油成分的研究

用水蒸气蒸馏法从荆条中提取挥发油,用气相色谱-质谱联用技术对挥发油成分进行分离和结构鉴定。从荆条籽的挥发油中鉴定出了51个组分,用气相色谱面积归一法确定了各个组分的相对百分含量,其挥发油的主要组成是β-榄香烯(27.98%),芳樟醇(12.39%),贝壳杉烯(12.00%),δ-榄香烯(10.54%),乙酯异冰片脂(8.98%)等。     

Fire mediated edge effects in bayhead tree islands

Question: The role edges play in mediating the effects of disturbance is unclear. Bayhead tree islands, which experience above‐ and belowground fire, contain trees that recover from disturbance by seed (Pinus elliottii var. densa) and by sprouting (Gordonia lasianthus). How does distance‐to‐edge affect survival and post‐fire response of trees with these contrasting life‐history strategies? Location: Two bayhead tree islands at Archbold Biological Station, central Florida, North America. Methods: Stem diameter, depth of peat smoldering, char height, resprouting status, and location were recorded for all Pinus and Gordonia stems ≥8 cm. Distance to the edge of the tree island was quantified using GIS. Results: The focal species showed contrasting patterns of survival across the edge‐to‐interior gradient that reflected gradients of fire severity. Survival of Gordonia was lowest in the bayhead interior where peat smolder was deepest. Conversely, survival of Pinus was lowest near the edges where char heights were greatest. The distinct types of Gordonia resprouting (crown versus basal) also showed spatially contrasting patterns. Basal resprouting dominated near the edges and was positively influenced by char height, while crown resprouting was nearly constant across the edge‐to‐interior gradient and was negatively influenced by char height. Conclusions: The spatial patterns of tree survival and resprouting observed are likely due to gradients in intensity of peat smoldering and aboveground burning, coupled with differential susceptibility to these two types of fire. Despite the rarity of fire in wetland tree islands (compared to uplands) it may play an important role in structuring the spatial distribution of trees.     

小麦族中间鹅观草不同居群的形态多样性分析

小麦族（Triticeae）植物的野外调查、收集通常是以形态学为依据的。为了探讨小麦族植物在野外调查、收集的科学取样策略,本项研究以小麦族具有自花授粉习性的3个中间鹅观草（Roegneria sinica. var. media Keng）居群、每个居群30个单株为材料,对11个形态学性状的多样性进行了分析。结果表明,3个居群的总遗传多样性指数为1.991,遗传多样性主要集中于居群内（91.76%）,而居群间的遗传变异（8.24%）相对较小;不同取样梯度下的遗传多样性指数随单株取样数目的增加呈现增大趋势,但当取样数目达到18株时,遗传多样性指数达到最高值。上述结果说明,对于小麦族自花授粉植物野外调查、收集时,应以居群为单位,而且每一居群至少应调查、收集18个单株,才能代表居群的遗传多样性。以形态学为依据的取样策略的建立,对于指导野外调查、收集具有现实意义。     

无绿藻的表型及其分子生物学鉴定

目的通过表型及分子生物学方法,正确鉴定无绿藻及其变种。方法回顾、总结与分析无绿藻的形态与结构、表型特征及生理生化及分子生物学鉴定、对常见抗真菌药物的敏感性及组织病理学特征。结果通过表型及分子生物学技术成功鉴定2株来自脑脊液及淋巴结的无绿藻:中型无绿藻碳水化合物变种及中型无绿藻波多黎各变种,并展示相关照片。结论无绿藻病的症状尚无特异性,其诊断主要依靠真菌学检查。标本的直接镜检、真菌培养及组织病理检查是主要手段。对无绿藻菌种的鉴定除了菌落形态、镜下结构外(含内孢子的孢子囊是无绿藻属的重要特征),糖类、醇类的同化利用,温度试验,结合分子生物学鉴定将有助于菌种的鉴定。     

内蒙古大兴安岭白头翁属一新变种

描述了毛茛科白头翁属一新变种——裂萼细叶白头翁(Pulsatilla turczaninovii Kryl.et Serg.var.fissasepalum)。本变种与原变种细叶白头翁的主要区别为,外轮萼片有不等深度、不等大小的二深裂,中裂片最大,两侧裂片大小不等。     

纪伊潮菊离体保存及其遗传稳定性分析

通过在基本培养基中添加不同浓度配比的蔗糖和矮壮素(CCC)对纪伊潮菊(Chrysanthemum shiwogiku var.kinokuniense)离体保存的影响进行研究,并对保存材料再生后代的遗传稳定性进行分子标记鉴定与分析.结果表明:在(23±2)℃、2 000～3 000 lx光照强度、12 h/d的光照培养条件下,MS+0.5 mg·L~(-1) BA+0.1 mg·L~(-1) NAA+琼脂6.5 g·L~(-1)培养基中添加30 g·L~(-1)蔗糖和1 500～2 000 mg·L~(-1)的CCC能够保存试管苗12个月,存活率为92.86%~96.43%,且恢复生长后试管苗长势良好,其再生后代的形态特征、过氧化物酶(POD)活性和ISSR分子标记扩增图谱与对照株无差异.     

新疆赖草属(Poaceae)两个新变种

报道了新疆赖草属2个新变种,即北疆赖草(Leymus tianschanicus (Drob.) Tzvel. var. borealus L. B. Cai)和短穗赖草(Leymus angustus (Trin.) Pilger var. brevistachyus L. B. Cai)。