拟穴青蟹GRP78基因的克隆与应激表达

葡萄糖调节蛋白78(glucose regulated protein 78 ku,GRP78)是热休克蛋白70家族成员之一,在调节蛋白质折叠和维持内质网稳态过程中起着分子伴 侣作用。采用RT-PCR、RACE等技术,首次从拟穴青蟹获得了GRP78的cDNA全长序列。该序列全长2 284 bp,开放阅读框(ORF)为1 962 bp,编码653个氨基酸残基。 同源分析显示,该基因编码的蛋白含有HSP70家族的签名序列,C末端为内质网蛋白滞留信号KDEL,与其他物种具有很高相似性。实时荧光定量PCR结果表明,GRP78 基因在拟穴青蟹多个组织中均有表达。第一期仔蟹在不同的温度和盐度下暴露12 h后,GRP78基因表达量随环境温度升高而增加;在高盐(30)条件下GRP78表达量 较高,进而推测拟穴青蟹GRP78参与蛋白质折叠和环境胁迫的应答。     

Einfluss selektiver mechanischer Fruchtbehangsregulierung auf Ethylensynthese als Stressindikator sowie Ertrag und Fruchtqualität bei Kernobst

The overall objective of this work was to improve fruit quality, break alternate bearing and reduce hand thinning using fewer chemicals in fruit crops. A device was constructed for mechanical thinning, which consisted of three independent horizontal rotors with ropes and freely adjustable angles on a frame, mounted on a front three point hitch and powered by the tractor hydraulics. This can be adapted to any fruit tree trained as spindle, Solaxe, (tall) vertical axis or fruit wall (le mur fruitier) irrespective of rootstock employed. Rotor speed varied from 300 to 460?rpm at either 5 or 7.5?km/h tractor speed. Eight-year-old or twelve-old apple trees cvs. ‘Gala’ and ‘Golden Delicious’ were mechanically thinned in 2007 between pink bud and full bloom (flower bud stages 6–8 or F1–F2) near Bonn, Germany; non-thinned and hand-thinned apple trees of the same block and variety served as control. Mechanically thinned flowering branches showed a similar amount of ethylene efflux (0.4–0.6?ppm C₂H₄/branch) as non-thinned flower branches, preventing potentially unexpected subsequent fruit drop, except for those removed by the rotors. The impact of the horizontal rotors on the branches was from the upper side and removed excessive flowers right to the tree trunk viz. the centre of the tree canopy, where fruits of lesser quality are expected leaving 2–3 flowers per cluster. Leaf damage was less than??10%, even at the fast rotor speed of 420?rpm, which was associated with negligible wood injury. Mechanical thinning induced firmer and sweeter fruit, i.e. tastier apples with longer shelf life, relative to control fruit from non-thinned apple trees. The greatest efficacy in terms of final fruit quality in the grading/sorting was achieved by a rotor speed of 360?rpm at a tractor speed of 5?km/h: Fruit mass increased by up to 20?g and the proportion of fruit larger than 70–75?mm by 10–30% compared with the fruit from non-thinned trees. Mechanical thinning with this newly constructed device led to a 10–20% reduction in yield, but increased returns due to better fruit size and colouration in apple with the potential to overcome alternate bearing.     

Molecular identification and genetic analysis for 24 turf-type Cynodon cultivars by Sequence-Related Amplified Polymorphism markers

Bermudagrass (Cynodon ssp.) germplasm is genetically diverse and widely distributed in the world. The study was conducted to identify and assess the molecular variation and relationship among 24 cultivars developed in China, Australia and the USA. Sequence-Related Amplified Polymorphism (SRAP) was applied to cultivars identification in this study for the first time. Thirty of the 90 SRAP primer combinations generated a total of 274 clearly bands encompassing 249 (91%) polymorphic. Each bermudagrass cultivar has its unique binary code and can be distinguished from the others. Three distinct clusters were obtained by unweighted pair-group method with arithmetic averages based on the polymorphic markers. Coefficients of genetic distance among the genotypes ranged from 0.57 to 0.97. The results demonstrated that SRAP marker is a stable molecular marker technique for the identification of bermudagrass cultivars and their genetic relationships.     

白菜亚硝酸还原酶基因BcNiR的克隆及表达分析

 以白菜品种‘苏州青’自交系叶片cDNA为模板, 采用RT-PCR、3′RACE和5′RACE技术, 获得了编码亚硝酸还原酶基因(NiR) 的cDNA全序列1 852 bp, 包含有1 749 bp的开放阅读框, 编码583个氨基酸, 命名为BcNiR。所推导的氨基酸序列与拟南芥NiR1、烟草nii2编码的氨基酸序列具有较高同源性, 分别为83%和76%。生物信息学分析表明, BcNiR具有完整的NiR蛋白结构, 含血红素蛋白β - 化合物区域, 一个明显的西罗血红素siroheme结合位点和4Fe-4S区域, 可以在SWiSS-MODEL数据库中搜索到与之相近的三维结构。RT-PCR结果显示, 该基因在叶片中的表达量远远高于根系, 在以0、10、20、30、40 mmol·L ^{- 1}硝态氮各分别处理0、2、4、6、8、12 h的试验中, 30 mmol·L ^-1处理4 h可使BcNiR的表达量达到最大; 5和10 mmol·L ^{- 1}铵态氮处理试验表明, 高浓度的铵抑制该基因的表达。     

利用DAD1反义片段转化创建菜薹可调控雄性不育材料

 菜薹因为没有好的雄性不育材料或自交不亲和系,至今尚无一代杂种用于生产,根据拟南芥及白菜型油菜的花药不开裂基因DAD1的保守序列设计引物,扩增菜薹的DAD1基因片段（DAD1F）,构建反义DAD1F植物表达载体,用农杆菌介导法转化菜薹,对转基因植株进行分子检测,鉴定其雄性不育性并进行育性恢复试验.克隆得到的菜薹的DAD1基因片段大小为678 bp,命名为BrcpDAD1F,其序列与拟南芥和白菜型油菜的DAD1高度同源,同源率分别为88％和99％;共得到了12株转基因植株,有6株在转录水平上得到表达,表现为雄性不育,花器官畸形,花粉活力低,萌发率不到10％,且开花后不能结角果或结空角果,或者得到极少种子但种子不萌发;用对照的花粉给转基因植株授粉可使其正常结实.以500 μmol·L^-1茉莉酸甲酯处理可使其雄性不育得到恢复,花粉可以在柱头和培养基上萌发,具有受精能力。T₁代可育株与不育株的比例都呈1:3分离, T₂代不同株系的育性分离比例不同,有些株系继续呈1:3的分离,有些株系全是可育株或全是不育株,说明反义抑制呈单基因稳定遗传。     

茄子离体叶片光系统Ⅱ的冷激胁迫效应及热力学分析

 以耐冷性较强的短把黑圆茄和耐热性较强的黑帅圆茄为试材,经冷激胁迫后采用植物效率仪PEA,进行快速叶绿素荧光参数测定,并利用最大光化学效率Fv/F m和活性中心RC/CSo的标准状态变性自由能变△GD进行PSⅡ冷稳定性的热力学分析。随着冷激胁迫温度的降低和时间的延长,PSⅡ的Fv/Fm、实际光化学效率ΔF/Fm′、RC/CSo呈"S"型下降趋势;单位反应中心DIo/RC和非光化学猝灭qN呈"S"型上升趋势;综合分析反映出冷激胁迫下PSⅡ反应中心的可逆失活和依赖于叶黄素循环的热耗散的双重机制在保护PSⅡ防止光抑制中起到重要作用。另外,通过两品种间PSⅡ行为差异,引入了两组新的参数作为便于直观比较冷胁迫的评价指标。黑帅圆茄Fv/Fm、ΔF/Fm＇的半衰温度T1/2和t1/2值分别高于短把黑圆茄1.4 ℃和0.8 ℃。茄子叶片Fv/Fm和RC/CSo的△GD随着冷激胁迫温度的降低而减少呈瀑布型的下降趋势。黑帅圆茄的△GD值和变性中点温度Tm都低于短把黑圆茄。它们均反映出黑帅圆茄的耐冷性低于短把黑圆茄。     

苹果园春季土施尿素的利用及其在土壤中的累积

 以7年生嘎拉苹果/平邑甜茶为试材, 利用¹⁵N示踪技术, 研究了苹果生产体系中氮素年周期 的利用、残留、损失及在土壤中的迁移动态。结果表明, 对春季土施尿素的利用率盛花期较低, 为11.38% , 至采收后达到最高。土壤氮素残留率盛花期最高, 为57.10% , 果实采后残留率最低。年周期中各土层氮素残留量不同, 盛花期20～40 cm土层残留量最高, 采收后最低, 0～20 cm土层在新梢旺长期残留量最高, 果实成熟期最低, 而40～60 cm土层在果实膨大期残留量达到最高, 在采收后0～20 cm土层残留有所增加, 其余各土层残留量均达到最低值。氮素损失与土壤残留呈相反的变化趋势, 氮素损失率在盛花期最低为31.53% , 随物候期的推迟逐渐升高, 在果实采后高达58.40%     

‘岩溪晚芦’椪柑果皮与果肉差减cDNA文库的构建及初步分析

 以‘岩溪晚芦’(Citrus reticulata Blanco)成熟果皮和果肉为材料,采用抑制性差减杂交技术,成功构建了果皮（tester）与果肉（driver）的差减cDNA文库。结果显示：cDNA克隆外源插入片段大小介于100～500 bp之间。成功对411个克隆进行了测序,测序结果经与NCBI基因库中序列进行比较,377个EST 找到了同源序列,它们分属于126个基因,涉及到与色素合成、能量代谢、初级代谢、次生代谢、抗逆防御、蛋白代谢、信号转导、香味形成、果皮软化（衰老）、精油合成、抗氧化代谢等代谢途径和生理生化过程。还有一些基因涉及到三价铁螯合物、反转录转座子gag蛋白、查尔酮合成、硼离子转运、儿茶酚氧位甲基转移酶等功能。功能已知且没有重复的EST共有63个,占总数的49.2%。有5个基因共62个克隆虽在Blast搜索时找到了较高的同源性序列,但功能目前尚未明确,有待进一步深入分析。另有34个基因未搜索到同源序列,可能为新发现的基因。     

The concept of habitat specificity revisited

Habitat specificity indices reflect richness (α) and/or distinctiveness (β) components of diversity. The latter may be defined by α and γ (landscape) diversity in two alternative ways: multiplicatively () and additively (). We demonstrate that the original habitat specificity concept of Wagner and Edwards (Landscape Ecol 16:121–131, 2001) consists of three independent components: core habitat specificity (uniqueness of the species composition), patch area and patch species richness. We describe habitat specificity as a family of indices that may include either area or richness components, or none or both, and open for use of different types of mean in calculation of core habitat specificity. Core habitat specificity is a beta diversity measure: the effective number of completely distinct communities in the landscape. Habitat specificity weighted by species number is a gamma diversity measure: the effective number of species that a patch contributes to landscape richness. We compared 12 habitat specificity indices by theoretical reasoning and by use of field data (vascular plant species in SE Norwegian agricultural landscapes). Habitat specificity indices are strongly influenced by weights for patch area and patch species richness, and the relative contribution of rare vs. common species (type of mean). The relevance of properties emphasized by each habitat specificity index for evaluation of patches in a biodiversity context is discussed. Core habitat specificity is emphasized as an ecologically interpretable measure that specifically addresses patch uniqueness while habitat specificity weighted by species number combines species richness and species composition in ways relevant for conservation biological assessment. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Regeneration of adventitious shoots from mature stored cotyledons of Japanese plum (Prunus salicina Lind1)

Adventitious shoot regeneration from mature stored cotyledons of Japanese plum (Prunus salicina Lind1) was achieved in vitro. The influences of the presence and absence of the light, different concentrations of thidiazuron (TDZ) and benzyladenine (BA) in the culture media, TDZ pretreatments and different basal salts on shoot regeneration were evaluated. TDZ was more effective in inducing shoot regeneration from mature stored cotyledons than BA. Dark incubation significantly increased the regeneration frequencies. Quoirin/Lepoivre (QL) basal salts stimulated shoot regeneration more than woody plant (WPM) or B5 salts did. The frequency of adventitious shoot formation varied among the varieties and the regeneration ability appeared to be genotype depended. The frequency of regeneration under the optimum tested conditions for ‘Bruce’, ‘Shiro’, ‘Redheart’, ‘Gladstone’ and ‘Early Golden’ cotyledons were 66.7%, 46.7%, 43.3%, 26.7% and 6.7%, respectively.