Sequence analysis and functional characterization of the promoter of the PiceaglaucaCinnamylAlcoholDehydrogenase gene in transgenic white spruce plants

The enzyme Cinnamyl Alcohol Dehydrogenase (CAD) catalyses the last step of lignin monomer synthesis, and is considered as a molecular marker of cell wall lignification in different plants species. Here, we report the isolation and analysis of 5′ flanking genomic DNA regions upstream to the CAD gene, from two conifers, i.e. white spruce (Picea glauca (Moench) Voss) and loblolly pine (Pinus taeda L.). Sequence comparisons with available CAD gene promoters from angiosperms highlighted the conservation of cis-elements matching MYB, WRKY and bHLH binding sites. Functional characterization of the P. glauca CAD promoter used P. glauca seedlings stably transformed with a DNA fragment of 1,163 base pairs (PgCAD) fused to the β-glucuronidase (GUS) gene. Histochemical observations of different vegetative organs of the transgenic trees showed that this sequence was sufficient to drive GUS expression in lignifying tissues, and more specifically in differentiating xylem cells. Quantitative RT-PCR experiments also indicated that the native CAD gene was preferentially expressed in differentiating xylem both in stems and roots. In addition, GUS expression driven by the PgCAD promoter was wound-inducible which was consistent with the accumulation of CAD mRNA in response to jasmonate application and mechanical wounding. The spruce CAD promoter represents a valuable tool for research and biotechnology applications related to xylem and wood. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Cold-regulated proteins with potent antifreeze and cryoprotective activities in spruces (Picea spp.)

Antifreeze proteins (AFPs) were obtained from intercellular spaces of spruce needles Picea abies (L.) Karst. and Picea pungens Engelm. by vacuum infiltration with ascorbic acid, followed by centrifugation to recover the infiltrate. As shown by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE), apoplastic proteins are accumulated in these spruce species as a group of 5–9 polypeptide bands. These proteins have a molecular mass of 7–80 kDa. The spruce AFPs have the ability to modify the growth of ice and thermal hysteresis, TH, caused by these AFPs was close to 2.0 °C at a concentration of 400 μg/ml. The antifreeze activity of proteins from these winter-hardy coniferous species showed a positive correlation with the concentration of proteins after cold acclimation of needle tissues. Apoplastic proteins from winter spruce needles exhibited antifreeze activity, whereas no such activity was observed in extracts from summer needles. When we examined the possible role of spruce AFPs in cryoprotection, we found that lactate dehydrogenase, LDH, activity was higher after freezing in the presence of AFPs compared with bovine serum albumin. Amino-terminal sequence comparisons indicated that a 27-kDa protein from both P. abies and P. pungens was similar to some pathogenesis-related proteins namely chitinases, also from conifer species. These results show that spruces produce AFPs that are secreted into the apoplast of needles. The accumulation of AFPs in extracellular spaces caused by seasonal cold acclimation during winter indicates that these proteins may play a role in the acquisition of freezing tolerance of needle cells in coniferous species.     

杜鹃花TPS基因家族鉴定及与萜类物质代谢的关系分析

萜烯合成酶（terpene synthase,TPS）能催化不同的前体物质生成不同的萜类化合物,是合成萜类物质的关键酶。为探究杜鹃花TPS基因家族成员在萜类物质代谢过程中的表达模式,本文基于杜鹃花基因组数据库,利用生物信息学方法对杜鹃花TPS基因（TPS）进行家族成员鉴定;通过云锦杜鹃和诺娃杜鹃两种不同种高山杜鹃的转录组测序结果,结合qRT-PCR、顶空固相微萃取和气相色谱-质谱联用技术,分析两种杜鹃不同发育时期花瓣中TPS家族成员表达水平和代谢物含量变化关系。结果表明,从杜鹃花基因组数据库中共鉴定获得47个RsTPS成员,RsTPS家族成员长度在591-2 634 bp之间,含有3-12个外显子不等,编码196-877个氨基酸;RsTPS家族成员主要分布在叶绿体和细胞质;系统进化分析结果显示RsTPS基因分为5个亚组。通过分析转录组数据得到7个功能注释为TPS的基因家族成员,发现TPS1、TPS10、TPS12和TPS13的表达量在4个时期中呈现出先上升,到盛开期达到顶峰后再下降的趋势。对基因表达量变化与萜类物质含量变化进行相关性分析,发现TPS1、TPS4、TPS9、TPS10、TPS12和TPS13表达量与云锦杜鹃不同时期花瓣中萜类物质含量变化呈显著性正相关,推测这6个基因家族成员可能是参与云锦杜鹃花香调控的关键基因。     

Food consumption and utilization by 6th-instar larvae of spruce budworm, Choristoneura fumiferana: a comparison on three Picea (spruce) species

The effects of tree species, shoot age, and sex of larvae on food consumption and utilization were investigated in a factorial experiment. Diets prepared from lyophilized and powdered current-year shoots, from two age classes (sample dates) of Picea glauca (Moench) Voss, Picea rubra, Sarg., and Picea mariana (Mill.) B.S.P., were evaluated for eight performance criteria. All factors had significant effects on all criteria except for tree species on assimilation efficiency, shoot age on development time, and sex on net growth efficiency. Eight of the possible 24 two-factor interactions and one of the possible eight three-factor interactions were significant. Food utilization efficiencies and relative growth rate were in the low and moderate characterization limits for immature arthropods; relative consumption rates were high. The higher performance values on Picea glauca were related to this species greater vulnerability to spruce budworm. Performance criteria of similar larvae on an artificial diet were compared with those on the foliage-diets.

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Résumé L'auteur a déterminé la quantité d'aliments utilisées, à 22±0,5 °C, par des larves de Tordeuse des bourgeons de l'épinette parvenues à leur sixième stade de développement qui s'étaient nourries d'un régime artificiel et d'aiguilles de l'année en cours lyophilisées et réduites en poudre, prélevées à deux dates différentes, à 10 jours d'intervalle, et provenant d'épinettes blanchés (Picea glauca [Moench] Voss), d'épinettes rouges (Picea rubra Sarg.) et d'épinettes noires (Picea mariana [Mill.] B.S.P.). Les femelles ont invariablement ingéré une quantité significativement plus importante de tous les régimes que les mâles, consommant 24% de plus d'aiguilles d'épinette noire et 73% de plus d'aiguilles d'épinette blanche.Les femelles prenaient en général plus de temps pour parvenir au stade de chrysalide et avaient un taux d'utilisation des aliments significativement plus élevé. L'efficacité nette de la croissance chez les deux sexes ne présentait toutefois aucune différence significative. La performance des larves ayant ingéré des aiguilles provenant de deux classes d'âge différait de façon significative mais non pas uniforme d'un régime à l'autre: les aiguilles d'épinette blanche plus récentes donnaient une performance généralement plus élevée contrairement aux aiguilles plus jeunes d'épinette rouge et d'épinette noire. Les larves ingéraient une quantité significativement plus élevée, avaient un taux de consommation et de croissance supérieur et augmentaient davantage leur biomasse avec des aiguilles d'épinette blanche. L'efficacité d'utilisation des aliments et le taux de croissance relatif étaient de faibles à modérément faibles pour un lépidoptère arboricole phyllophage; la vitesse d'ingestion relative était élevée. La qualité des aiguilles des trois essences d'épinettes était faible. Les aiguilles d'épinette blanche permettaient aux larves d'avoir une performance significativement plus élevée, performance mesurée à partir de plusieurs critères, alors que les aiguilles d'épinette rouge et d'épinette noire donnaient une performance égale. La supériorité des performances obtenues avec l'épinette blanche explique la plus grande vulnérabilité de cette essence à la Tordeuse des bourgeons de l'épinette.

     

Gene structure,phylogeny and expression profile of the sucrose synthase gene family in cacao (Theobroma cacao L.)

Journal of Genetics - In higher plants, sucrose synthase (Sus, EC 2.4.1.13) is widely considered as a key enzyme involved in sucrose metabolism. Although, several paralogous genes encoding...     

Rhodopechys obsoleta (desert finch): a pale ancestor of greenfinches ( Carduelis spp.) according to molecular phylogeny

The evolutionary history of three out of four birds traditionally classified into the genus Rhodopechys birds has been studied by comparing their mitochondrial cytochrome b DNA sequence with that of greenfinches and other genus Carduelis finches. The desert finch (Rhodopechys obsoleta) or a sister extinct species seems to have existed about 6 million years ago in Asian and perhaps African desert-like areas. This bird has no molecular relationship with other Rhodopechys birds and seems to have given rise to the greenfinches radiation, probably by allopatry of marginal or isolated groups; the latter would have evolved to green plumage colours and more simple song modulations (i.e., greenfinches). The possible role of assortative mating and the newly postulated acquired phenotypic characters in greenfinches speciation are discussed.     

Genome-wide analysis of the terpene synthase gene family in physic nut (<Emphasis Type="Italic">Jatropha curcas</Emphasis> L.) and functional identification of six terpene synthases

Plant terpenes constitute a large class of compounds and have numerous biological roles as either primary or secondary metabolites. Terpene synthases (TPSs) play key roles on catalyzing the formation of different terpenes; they are divided into seven subfamilies based on sequence relatedness (TPSa–h). TPS-a proteins catalyze the formation of sesquiterpenes and diterpenes in plants. Physic nut (Jatropha curcas L.) is an attractive biofuel tree, but its seeds contain diterpene derivatives, which make them inedible for animals. In this study, 59 putative TPS genes (JcTPS01 to JcTPS59) were identified in the physic nut genome, and 26 belong to the TPS-a subfamily. Eight among the 26 TPS-a genes showed expression in developing seeds of physic nut in the present study. After heterologous expression of these eight genes in Escherichia coli and in vitro enzyme assays, six were shown to have TPS activities. Two (JcTPS09 and JcTPS11) catalyzed the production of diterpene casbene, which was consistent with earlier findings. The other four (JcTPS02, 23, 55, and 56) catalyzed the production of sesquiterpenes. These results may facilitate the efforts for identifying TPS genes involving the physic nut terpene synthesize.     

The cinnamyl alcohol dehydrogenase gene family in Populus: phylogeny, organization, and expression

Background  

Lignin is a phenolic heteropolymer in secondary cell walls that plays a major role in the development of plants and their defense against pathogens. The biosynthesis of monolignols, which represent the main component of lignin involves many enzymes. The cinnamyl alcohol dehydrogenase (CAD) is a key enzyme in lignin biosynthesis as it catalyzes the final step in the synthesis of monolignols. The CAD gene family has been studied in Arabidopsis thaliana, Oryza sativa and partially in Populus. This is the first comprehensive study on the CAD gene family in woody plants including genome organization, gene structure, phylogeny across land plant lineages, and expression profiling in Populus.     

Nitrate removal and DO levels in batch wetland mesocosms: Cattail (Typha spp.) versus bulrush (Scirpus spp.)

Nitrate removal rates and dissolved oxygen (DO) levels were evaluated in small batch-mode wetland mesocosms with two different plant species, cattail (Typha spp.) and bulrush (Scirpus spp.), and associated mineral-dominated sediment collected from a mature treatment wetland. Nitrate loss in both cattail and bulrush mesocosms was first-order in nature. First-order volumetric rate constants (k_V) were 0.30 d⁻¹ for cattail and 0.21 d⁻¹ for bulrush and rates of nitrate loss were significantly different between plant treatments (p < 0.005). On an areal basis, maximum rates of nitrate removal were around 500 mg N/(m² d) early in the experiment when nitrate levels were high (> 15 mg N/L). Areal removal rates were on average 25% higher in cattail versus bulrush mesocosms. DO in mesocosm water was significantly higher in bulrush versus cattail (p < 0.001). DO in bulrush generally ranged between 0.5 and 2 mg/L, while DO in cattail mesocosms was consistently below 0.3 mg/L. Based on cumulative frequency analysis, DO exceeded 1 mg/L around 50% of the time in bulrush, but only 2% of the time in cattail. DO in bulrush exhibited a statistically significant diel cycle with DO peaks in the late afternoon and DO minimums in the early morning hours. Difference in nitrate removal rates between wetland plant treatments may have been due to differing plant carbon quality. Cattail litter, which has been shown in other studies to exhibit superior biodegradability, may have enhanced biological denitrification by fueling heterotrophic microbial activity, which in turn may have depressed DO levels, a prerequisite for denitrification. Our results show that the cattail is more effective than bulrush for treating nitrate-dominant wastewaters.     

茄子TCP基因家族全基因组的鉴定与分析

TCP (teosinte branched1/cincinnata/proliferating cell factor)转录因子是植物特有转录因子家族,在植物整个生长发育过程中都有着很重要作用。目前,在茄子(Solanum melongena L.)中还没有关于TCP转录因子的相关报道。本研究利用生物信息学方法在茄子基因组数据库中鉴定出分布于11条染色体上的29个茄子TCP家族基因(eggplant TCP,SmTCP)。研究结果显示,该家族所有成员均含有编码TCP保守结构域的序列。这些成员氨基酸长度范围为201–538 aa,外显子数为1或2。亚细胞定位显示,有3个SmTCP (SmTCP02/03/21)蛋白位于细胞质中,其他SmTCP蛋白都位于细胞核中。系统发育树和序列特征分析均将29个SmTCP基因分成ClassⅠ(PCF)和ClassⅡ(CIN和CYC/TB1)两大类。共线性分析发现,17对(21个)SmTCP基因存在共线性关系,且这些存在共线性关系的基因都属于片段复制。基因表达模式分析显示,29个SmTCP基因家族成员在15个组织或器官中都有表达,但是不同成员的表达模式存在差异,其中CIN亚家族的4个基因(SmTCP18/19/20/25)在3个不同生长期的叶片中均较高表达。SmTCP启动子区域的顺式作用元件分析共发现4类顺式作用元件。本研究从多个角度分析了SmTCP基因分子基础,研究了TCP转录因子对茄子生长发育的影响,为茄子分子育种提供理论参考。