Seed dormancy patterns in Epipactis palustris (Orchidaceae): Requirements for germination and establishment of mycorrhiza

Some terrestrial orchid species, including Epipactis palustris (L.) Crantz, are considered extremely difficult to germinate and cultivate in vitro. Observations of orchids germinating in nature are very few, and the timing and requirements for seedling establishment are unknown for most species. Seeds of E. palustris were incubated in vitro with an appropriate fungus, but germination was poor unless several other conditions were also met: scarification of the testa in Ca(OCL)₂, an initial incubation for several weeks at 27°C, and a subsequent cold stratification for 8–12 weeks at 4–8°C, With these pretreatments, germnation responses exceeded 50% after incubation for 4 weeks at 20°C. Healthy protocorms with normal organ development were only produced by symbiotic culture following this lengthy seed preparation. The findings suggest that under natural conditions the seeds need some after-ripening, and the testa needs to be partially decomposed before germination. The requirement for chilling suggests that germination of seeds in situ occurs in spring.     

人血小板生成素全长分子在酵母系统中的分泌表达及活性分析

外源基因可通过整合型载体被整合到毕赤酵母（Ｐｉｃｈｉａｐａｓｔｏｒｉｓ）染色体上,获得遗传性稳定分泌表达株．利用酵母信号肽ＭＦα,对该信号肽蛋白酶识别位点的相关序列进行重新设计,使天然人ＴＰＯ成熟肽在毕赤酵母系统中分泌表达成功．表达产物经Ｗｅｓｔｅｒｎ－ｂｌｏｔ进行分析,分子量约为６６ｋＤ处蛋白条带可被ＴＰＯ抗体识别;表达量约为０．１ｇ／Ｌ;Ｎ端氨基酸序列分析结果与设计的一致;表达产物对小鼠骨髓细胞形成巨核细胞集落形成单位（ｍｅｇａｋａｒｙ－ｏｃｙｔｅｃｏｌｏｎｙｆｏｒｍｉｎｇｕｎｉｔ,ＣＦＵ－Ｍｅｇ）具有明显的刺激作用．     

人肝金属硫蛋白-I_A基因在鱼腥藻中的克隆与表达

将人工合成的人肝金属硫蛋白（ｍｅｔａｌｏｔｈｉｏｎｅｉｎ,简称ＭＴ）－ＩＡ基因插入至中间载体ｐＲＬ－４３９上强启动子ｐｓｂＡ后,再将其与穿梭载体ｐＫＴ－２１０相连,得到大肠杆菌－蓝藻穿梭表达载体ｐＫＴ－ＭＴ,用三亲接合转移法将ｐＫＴ－ＭＴ转入丝状体蓝藻－鱼腥藻７１２０,经链霉素筛选,得到了稳定的转人肝ＭＴ－ＩＡ基因鱼腥藻．纯化单藻落,液体扩大培养．从鱼腥藻中提取的质粒经Ｓｏｕｔｈｅｒｎ印迹分析,确定人肝ＭＴ－ＩＡ基因已转入鱼腥藻７１２０中,Ｗｅｓｔｅｒｎ印迹分析表明,金属硫蛋白在转人肝ＭＴ－ＩＡ基因鱼腥藻中得到了表达．经原子吸收光谱法测定表达量约为７００μｇＭＴ／ｇ鲜藻,重金属耐受性实验表明,得到了能耐受重金属－镉的转人肝ＭＴ－ＩＡ基因鱼腥藻,它将在清除水域中重金属污染和医药研究方面发挥重要作用．     

How does a wetland plant respond to increasing temperature along a latitudinal gradient?

Global warming affects plant fitness through changes in functional traits and thereby ecosystem function. Wetlands are declining worldwide, and hence, ecosystem functions linked to wetlands are threatened. We use Caltha palustris “a common wetland plant” to study whether warming affects growth and reproduction differently depending on origin of source population, potentially affecting phenotypic response to local climate. We conducted a 2‐year in situ temperature manipulation experiment using clone pairs of C. palustris in four regions, along a 1300‐km latitudinal gradient of Sweden. Open‐top chambers were used to passively increase temperature, paired with controls. Growth and reproductive traits were measured from 320 plants (four regions × five sites × two treatments × eight plants) over two consecutive seasons to assess the effect of warming over time. We found that warming increased plant height, leaf area, number of leaves, and roots. High‐latitude populations responded more strongly to warming than low‐latitude populations, especially by increasing leaf area. Warming increased number of flowers in general, but only in the second year, while number of fruits increased in low‐latitude populations the first year. Prolonged warming leads to an increase in both number of leaves and flowers over time. While reproduction shows varying and regional responses to warming, impacts on plant growth, especially in high‐latitude populations, have more profound effects. Such effects could lead to changes in plant community composition with increased abundance of fast‐growing plants with larger leaves and more clones, affecting plant competition and ecological functions such as decomposition and nutrient retention. Effects of warming were highly context dependent; thus, we encourage further use of warming experiments to predict changes in growth, reproduction, and community composition across wetland types and climate gradients targeting different plant forms.     

Genetic structure and AFLP variation of remnant populations in the rare plant Pedicularis palustris (Scrophulariaceae) and its relation to population size and reproductive components

We investigated plant reproduction in relation to genetic structure, population size, and habitat quality in 13 populations of the rare biennial plant Pedicularis palustris with 3-28500 flowering individuals. We used AFLP (amplified fragment length polymorphism) profiles to analyze genetic similarities among 129 individuals (3-15 per population). In a cluster analysis of genetic similarities most individuals (67%) were arranged in population-specific clusters. Analysis of molecular variance indicated significant genetic differentiation among populations and among and within subpopulations (P < 0.001). Gene flow (N(e) m) was low (0.298). On average, plants produced 55 capsules, 17 seeds per fruit, and 42 seedlings in the following growing season. The number of seeds per capsule was independent of population size and of genetic variability. In contrast, the number of capsules per plant (P < 0.05) and the number of seedlings per plant (P < 0.05) were positively correlated with population size. The relation between population size and the number of seeds per plant was not significant (P = 0.075). The number of capsules and of seeds and seedlings per plant (P < 0.01) were positively correlated with genetic variability. Genetic variability was independent of actual population size, suggesting that historical population processes have to be taken into account, too. Stepwise multiple regressions revealed additional significant relationships of habitat parameters (soil pH, C:N ratio), vegetation composition, and standing crop on reproductive components. We conclude that populations of P. palustris are genetically isolated and that reproductive success most likely is influenced by population size, genetic variability, and habitat quality. Management strategies such as moderate grazing, mowing, and artificial gene flow should endeavor to increase population size as well as genetic variation.     

Ultrastructural Study of Secondary Wall Formation in the Stem Fiber of Phyllostachys pubescens

Ultrastructural changes in secondary wall formation of Phyllostachys pubescens Mazel fiber were investigated with transmission electron microscopy. Fiber developed initially with the elongation of cells containing ribosomes, mitochondria and Golgi bodies in the dense cytoplasm. During the wall thickening, the number of rough endoplasmic reticulum and Golgi bodies increased apparently. There were two kinds of Golgi vesicles, together with the ones from endoplasmic reticulum formed transport vesicles. Many microtubules were arranged parallel to the long axis of the cell adjacent to the plasmalemma. Along with the further development of fiber, polylamellate structure of the secondary wall appeared, with concurrent agglutination of chromatin in the nucleus, swelling and disintegration of organelles, while cortical microtubules were still arranged neatly against the inner side of plasmalemma. Lomasomes could be observed between the wall and plasmalemma. The results indicated that the organelles, such as Golgi bodies together with small vesicles, rough endoplasmic reticulum and lomasomes, played the key role in the thickening and lignification of the secondary wall of bamboo fiber, though cortical microtubules were correlative with the process as well.