Programmed cell death during development of cowpea (Vigna unguiculata (L.) Walp.) seed coat

The seed coat develops primarily from maternal tissues and comprises multiple cell layers at maturity, providing a metabolically dynamic interface between the developing embryo and the environment during embryogenesis, dormancy and germination of seeds. Seed coat development involves dramatic cellular changes, and the aim of this research was to investigate the role of programmed cell death (PCD) events during the development of seed coats of cowpea [Vigna unguiculata (L.) Walp.]. We demonstrate that cells of the developing cowpea seed coats undergo a programme of autolytic cell death, detected as cellular morphological changes in nuclei, mitochondria, chloroplasts and vacuoles, DNA fragmentation and oligonucleosome accumulation in the cytoplasm, and loss of membrane viability. We show for the first time that classes 6 and 8 caspase‐like enzymes are active during seed coat development, and that these activities may be compartmentalized by translocation between vacuoles and cytoplasm during PCD events.     

Complete sequences of the rRNA genes of Drosophila melanogaster

In this, the first of three papers, we present the sequence of the ribosomal RNA (rRNA) genes of Drosophila melanogaster. The gene regions of D. melanogaster rDNA encode four individual rRNAs: 18S (1,995 nt), 5.8S (123 nt), 2S (30 nt), and 28S (3,945 nt). The ribosomal DNA (rDNA) repeat of D. melanogaster is AT rich (65.9% overall), with the spacers being particularly AT rich. Analysis of DNA simplicity reveals that, in contrast to the intergenic spacer (IGS) and the external transcribed spacer (ETS), most of the rRNA gene regions have been refractory to the action of slippage-like events, with the exception of the 28S rRNA gene expansion segments. It would seem that the 28S rRNA can accommodate the products of slippage-like events without loss of activity. In the following two papers we analyze the effects of sequence divergence on the evolution of (1) the 28S gene "expansion segments" and (2) the 28S and 18S rRNA secondary structures among eukaryotic species, respectively. Our detailed analyses reveal, in addition to unequal crossing-over, (1) the involvement of slippage and biased mutation in the evolution of the rDNA multigene family and (2) the molecular coevolution of both expansion segments and the nucleotides involved with compensatory changes required to maintain secondary structures of RNA.      

栲树不同生长发育阶段的枝系特征分析

对浙江天童国家森林公园内常绿阔叶优势树种栲树(Castanopsis fargesii)不同发育阶段植冠内的分枝式样特征进行了统计分析。结果表明：栲树在不同发育阶段的总体分枝率和逐步分枝率有显著变化,幼苗和幼树阶段的分枝率较低,而成株阶段的分枝率较高;幼树阶段的枝条长度、枝倾角和叶倾角明显大于幼苗和成株阶段,表现为明显的高生长对策;叶片配置在不同枝系上有较大差异,叶片主要集中于植冠内一级枝和二级枝上;叶片的大小从幼苗、幼树到成株阶段逐渐增大。研究结果表明栲树在生活史的不同生长发育阶段,分枝式样表现出一定的可塑性,反映了不同的适应对策。     

新的端粒酶活性抑制蛋白LPTS-L的制备

LPTS基因是利用定位候选克隆策略克隆的一个新的肝相关候选肿瘤抑制基因。LPTS基因编码一个全长为328氨基酸的蛋白质(LPTS-L),该蛋白具有抑制细胞端粒酶活性的功能。为了进一步研究LPTS-L蛋白的结构与功能,利用DNA重组技术,将LPTS-L的cDNA克隆到表达载体pET-24a中构建重组克隆pET-24-LPTS,并在大肠杆菌BL-21中进行融合表达,获得可溶形式的LPTS-L融合蛋白。采用Ni Sepharose4B柱亲和层析,可以获得纯度较高的蛋白,但不适合大量制备。通过设计引物去掉了pET-24a载体上的6×His tag将LPTS-L基因进行了非融合表达,然后采用磷酸纤维素P11阳离子交换层析纯化LPTS-L蛋白,纯度可达到55%。再经Sephadex G-100凝胶过滤,LPTS-L蛋白的纯度可达到80%。Western blot实验显示经纯化后的LPTS-L蛋白可与兔抗GST-LPTS-L的多抗发生特异性结合。采用TRAP法测定蛋白质活性,结果显示纯化得到的LPTS-L蛋白可抑制端粒酶的活性,与采用Ni Sepharose4B纯化获得的LPTS-L融合蛋白比较,其抑制效率基本一致。因此,所建立的技术可以有效地制备LPTS-L蛋白。     

Microsatellite loci for the stingless bee Melipona rufiventris (Hymenoptera: Apidae)

Eight microsatellite primers were developed from ISSR (intersimple sequence repeats) markers for the stingless bee Melipona rufiventris. These primers were tested in 20 M. rufiventris workers, representing a single population from Minas Gerais state. The number of alleles per locus ranged from 2 to 5 (mean = 2.63) and the observed and expected heterozygosity values ranged from 0.00 to 0.44 (mean = 0.20) and from 0.05 to 0.68 (mean = 0.31), respectively. Several loci were also polymorphic in M. quadrifasciata, M. bicolor, M. mandacaia and Partamona helleri and should prove useful in population studies of other stingless bees.     

Transpiration of trees and forest stands: short and long-term monitoring using sapflow methods

We show that sapflow is a useful tool for studies of water fluxes in forest ecosystems, because (i) it gives access to the spatial variability within a forest stand, (ii) it can be used even on steep slopes, and (iii) when combined with eddy correlation measurements over forests, it allows separation of individual tree transpiration from the total water loss of the stand. Moreover, sapflow techniques are quite easy to implement. Four sapflow techniques currently coexist, all based on heat diffusion in the xylem. We found a good agreement between three of these techniques. Most results presented here were obtained using the radial flow meter (Granier 1985). Tree sapflow is computed as sap flux density times sapwood area. To scale up from trees to a stand, measurements have to be made on a representative sample of trees. Thus, a number of trees in each circumference class is selected according to the fraction of sapwood they represent in the total sapwood area of the stand. The variability of sap flux density among trees is usually low (CV. 10–15%) in close stands of temperate coniferous or deciduous forests, but is much higher (35–50%) in a tropical rain forest. It also increases after thinning or during a dry spell. A set of 5–10 sapflow sensors usually provides an accurate estimate of stand transpiration. Transpiration measured on two dense spruce stands in the Vosges mountains (France) and one Scot's pine plantation in the Rhine valley (Germany) showed that maximum rate was related to stand LAI and to local climate. Preliminary results comparing the sapflow of a stand of Pinus banksiana to the transpiration of large branches, as part of the BOREAS programme in Saskachewan, Canada showed a similar trend. For modelling purposes, tree canopy conductance (g_c) was calculated from Penman-Monteith equation. In most experiments, calculated canopy conductance was dependent on global radiation (positive effect) and on vapour pressure deficit (negative effect) in the absence of other limiting factors. A comparison of the vapour pressure deficit response curves of g_c for several tree species and sites showed only small differences among spruce, oak and pine forests when including understorey. Tropical rainforests exhibited a similar behaviour.     

锁阳和肉苁蓉寄生方式的区别

锁阳和肉苁蓉都是中医药里重要的补益类药材,但由于过度采挖和采挖方式不当,目前它们的野生资源已濒临枯竭。肉苁蓉和锁阳分别是我国濒危和易危珍稀植物,研究二者寄生方式的特点与区别不仅可以促进锁阳和肉苁蓉的人工栽培,从而使野生药材得到一定的保护,而且对了解寄生植物在荒漠地区等极端严酷环境中的适应机制具有重要的生态学意义。该研究采用形态学观察结合常规石蜡切片法,对锁阳和肉苁蓉分别在各自寄主植物上的寄生方式进行了研究。结果表明：(1)锁阳的营养繁殖体在寄主植物根部呈串状分布,与寄主植物的连接方式属于非末端寄生;锁阳的吸器侵入寄主根系韧皮部和木质部的一部分区域,但是韧皮部和木质部大部分区域未被锁阳吸器占据,即有部分营养物质被锁阳“截取”。(2)肉苁蓉在其肉质茎基部长出新的芽体,与寄主植物的连接方式属于末端寄生;肉苁蓉的吸器侵入寄主根韧皮部和木质部全部区域。因此,锁阳寄生后,被寄生的寄主根依然能够向前生长,具有正常的功能;肉苁蓉寄生后,寄生点的寄主根失去根系的正常功能,成为一个为肉苁蓉生长发育提供营养物质的“输送通道( Transport channel)”。