中国樱桃CBF/DREB转录因子PpcCBF的克隆与表达分析

低温诱导中国樱桃(Prunus pseudocerasus)花芽休眠解除是一个非常复杂的过程。研究花芽响应低温的分子生物学机制将是揭示这一问题的关键。CBF/DREB转录因子在植物低温响应过程中发挥着重要作用。为此,本研究从"短柄"樱桃中克隆了一个CBF/DREB转录因子,命名为PpcC BF。生物信息学分析表明,Ppc CBF基因的开放阅读框为720 bp,编码239个氨基酸。Ppc CBF具有典型的CBF/DREB转录因子的结构特征,AP2结合域高度保守,聚类分析发现李属植物CBF/DREB转录因子亲缘关系较近。实时定量表达分析发现,Ppc CBF受低温诱导表达,且不依赖ABA。在花芽休眠解除过程中,随着低温积累而表达上调,休眠解除临界期之后下调表达,与花芽休眠解除具有极大相关性。     

植物冷驯化的分子机理研究进展

植物冷驯化是一个非常复杂的过程,包括植物将感受到的低温信号转变成生化信号,以激活冷诱导基因的启动子,刺激特定的mRNA的转录,并在特定的组织中合成冷驯化蛋白.冷驯化蛋白通过增强膜脂流动性和阻止胞间冰晶形成等方式,以保护细胞免受低温伤害.冷驯化基因的表达以转录后调控为主,也有转录调控.某些冷诱导基因也可受ABA或其他环境胁迫(如高温、干旱、高盐、脱水等)诱导表达.     

休眠期间油桃花芽碳水化合物代谢及其相关基因的表达变化

以十年生大田和三年生盆栽‘曙光’油桃花芽为材料,分别测定了其休眠期碳水化合物含量、糖代谢相关基因的季节性表达及低温处理下相关基因的表达变化,旨在探讨碳水化合物及低温与休眠的关系。结果表明：休眠期间可溶性糖（主要是蔗糖）含量逐渐增加,淀粉含量则呈相反趋势。糖代谢相关基因表达明显不同：腺苷二磷酸葡萄糖焦磷酸化酶基因口GPase）无明显变化;组氨酸H3基因（HisH3）和己糖激酶I基因（胱,）在进入内休眠前有明显上升,蔗糖合酶基因（SuSy）则与之相反;尿苷二磷酸葡萄糖焦磷酸化酶（UGPase）表达总体上呈上调趋势,在进入内休眠后稍有下调。表明进入内休眠后,依赖HKl的糖信号转导途径起重要作用。在4℃处理后,与细胞分裂有关的基因HisH3含量急剧升高,而后下降,说明细胞分裂的减少并不是休眠期间抑制生长的原因;UGPase表现出与内休眠期一致的变化趋势,说明对低温有一定的适应性。     

木本植物季节性休眠的分子机制

季节性休眠是多年生木本植物在生态和进化上的一种"权衡"机制,也是植物界多样性生存策略的组成部分.木本植物的休眠反应首先是Ca2+作为信号物质诱导CO/FT基因的表达,进而引起与休眠相关的DAM基因的表达;而低温可以诱导休眠的发生,冷诱导表达的基因CBF和COR在休眠期间也有表达;而与逆境相关的蛋白如脱水蛋白、抗冻蛋白、热激蛋白和热稳定蛋白等也与休眠反应有密切的关系.本文就与木本植物季节性休眠相关的分子机制进行了综述.     

低温诱导大百合鳞茎休眠解除与酚类代谢关系

大百合(Cardiocrinum giganteum)为多年生球根药食同源植物,其鳞茎具有典型的生理休眠特性,而低温是百合鳞茎解除休眠的重要环境因子。为揭示大百合鳞茎休眠解除的分子机制,该研究对4℃低温处理0、30和60d的鳞茎分别进行代谢组和转录组分析。结果表明,鳞茎休眠的解除与酚类物质的代谢相关,酚类物质的降解有利于解除休眠,其中苯丙氨酸解氨酶基因(PALs)在此过程中可能起主要作用。同时,bHLH、bZIP、MYB和MADS等转录因子家族成员均与酚类代谢物显著相关,且参与解除休眠。共表达分析证实PAL、CAD和POD是酚类代谢重要的调控基因,MYB4、MYB114和ICE1参与了酚类代谢调控网络,其中ICE1可能是连接温度信号和酚类代谢的关键因素。这些转录因子与酚类物质的共同作用可能对鳞茎打破休眠具有重要作用。     

不同低温需求量砂梨休眠相关基因PpMADS13的特征及表达模式

为探究福建主栽的不同低温需求量砂梨Pyrus pyrifolia品种中休眠相关基因PpMADS13-1和PpMADS13-3序列及表达特征,以‘黄花’、‘蜜雪’两品种休眠时期的花芽为材料,克隆PpMADS13-1和PpMADS13-3基因,利用荧光定量PCR技术分析基因在休眠时期的表达变化情况。结果显示,PpMADS13-1序列在砂梨两品种间较保守,而PpMADS13-3序列在两品种间保守性较差;‘黄花’梨PpMADS13-1hh和PpMADS13-3hh在休眠期间随着休眠的解除下调表达,‘蜜雪’梨PpMADS13-1mx和PpMADS13-3mx在休眠期间呈现“M”型表达趋势。PpMADS13-1和PpMADS13-3均对砂梨的花芽休眠解除起到调控作用,在不同低温需求量品种间PpMADS13-1和PpMADS13-3的休眠调控模式有所不同。     

高温和单氰胺对油桃休眠花芽呼吸代谢的影响

以3年生盆栽‘曙光’油桃为材料,研究油桃自然休眠过程中50℃高温和单氰胺对花芽呼吸代谢的影响.结果表明:高温和单氰胺均可以打破油桃的自然休眠,导致休眠花芽呼吸代谢显著下降,其呼吸代谢的衰减可持续数小时.主要呼吸途径三羧酸循环(TCA)和磷酸戊糖途径(PPP)的运行均受到影响.未经破眠处理的花芽TCA和PPP均呈衰减趋势,而高温和单氰胺诱导了早期呼吸衰减结束后PPP的迅速活化.高温还表现出对TCA恢复的诱导,而单氰胺在96 h内未表现出这种作用.在高温和单氰胺打破自然休眠的机制中,呼吸衰减和随后出现的PPP活化可能是重要的组成部分.     

水曲柳种子次生休眠的预防和解除

以解除休眠和经高温诱导产生次生休眠的水曲柳种子为材料,探讨预防和解除水曲柳种子次生休眠的方法。结果表明：GA3和乙烯利不能阻止已解除休眠水曲柳种子在25℃下萌发时诱导的次生休眠。干燥和短时间低温解除水曲柳种子次生休眠的效果不明显,较长时间（大于2周）的低温对解除水曲柳种子的次生休眠有一定的效果。综合来看,10^-3mol·L^-1的乙烯利或GA4＋7解除水曲柳种子次生休眠的效果较好。     

水孔蛋白在细胞延长、盐胁迫和光合作用中的作用

水孔蛋白属于一个高度保守的、能够进行跨生物膜水分运输的通道蛋白MIP家族。水孔蛋白作为膜水通道,在控制细胞和组织的水含量中扮演重要角色。本研究的重点是属于PIP亚家族的GhPIP1;2和属于TIP亚家族的γTIP1在植物细胞延长中的作用。使用特异基因探针的Northern杂交和实时荧光PCR技术证明GhPIP1;2和GhγTIP1主要在棉花纤维延长过程中显著表达,且最高表达量在开花后5d。在细胞延长过程中,GhPIP1;2和GhγTIP1表达显著,表明它们在促使水流迅速进入液泡这一过程中扮演重要角色。而且也研究了盐胁迫植物中钙离子对水孔蛋白的影响。分别或一起用NaCl或CaCl2处理原生质体或细胞质膜。结果发现在盐胁迫条件下,水渗透率值在原生质体和质膜颗粒中都下降了,同时PIP1水孔蛋白的含量也下降了,表明NaCl对水孔蛋白的功能和含量有抑制作用。同时也观察了Ca2+的两种不同的作用。感知胁迫的胞质中游离钙离子浓度的增加可能导致水孔蛋白的关闭。而过剩的钙离子将导致水孔蛋白的上游调控。同时实验已经证明大麦的一类水孔蛋白-HvPIP2;1有更高的水和CO2转移率。本研究的目标是确定负责转运水和CO2的关键水孔蛋白...     

植物耐冷性基因工程

温度决定物种的分布,同时还影响作物的产量和品质。植物耐冷的机制涉及到许多方面,包括膜脂组成的变化、可混溶溶质的积累、抗氧化酶活性的提高、低温相关基因的诱导表达等。由于植物的耐冷性状由多基因控制,采用传统的育种方法往往难以取得理想的结果,而植物基因工程技术的发展及应用则提供了另外可能的途径,可以通过转移耐冷性状形成的关键基因从而对植物进行改良。本文从膜脂组成、可混溶溶质、抗冻蛋白、抗氧化酶和诱导植物低温相关基因的转录因子等方面对植物耐冷性的基因工程研究进行了综述,以期为植物育种者和从事冷胁迫机制研究的工作者提供参考。     

桃芽自然休眠与两条主要电子传递途径变化的关系

花芽和叶芽总呼吸速率最低点均与自然休眠进程有关,第一个与自然休眠的起始时间相对应,最后一个则与自然休眠解除期相对应;细胞色素途径抑制剂氰化钾（KCN）对休眠芽的呼吸起部分抑制作用;抗氰呼吸抑制剂水杨基氧肟酸（salicylhydroxamic acid,SHAM）对总呼吸速率的效应随休眠进程而变化,休眠前期起促进作用,随休眠进程其促进作用逐渐减弱,从可调控休眠期（对外源措施敏感期）起转入抑制效应;KCN＋SHAM混合剂对总呼吸速率的效应与SHAM单独使用的效果相似,但其时总呼吸速率促进作用的起始点和结束点均较SHAM单独使用旱7d左右。     

几种化学药剂对‘NJ72’油桃休眠解除的影响

在进行果树温室栽培时,经常遇到萌芽率低、萌芽开花延迟、花器官发育差、座果率低的问题。本试验以‘NJ72’油桃为试材,观察了3种药剂对解除芽休眠的影响。结果表明,2%(NH2)2CS能提早花期,但存在药害现象。6% KNO3不能提早花期,并且花期不整齐,5% NH4NO3效果与6%KNO3类似。同时化学药剂处理促进花芽内H2O2的积累,抑制了过氧化氢酶(CAT)活性但促进了过氧化物酶(POD)活性,超氧物岐化酶(SOD)活性变化较小。化学药剂处理使花芽的呼吸速率增加,其中磷酸戊糖途径(PPP)代谢增加,糖酵解(EMP)降低,而三羧酸循环(TCA)代谢波动较小。葡萄糖_6_磷酸脱氢酶(G6PDH)活性在化学药剂处理时也增加。     

几种化学药剂对‘NJ72’油桃休眠解除的影响

在进行果树温室栽培时,经常遇到萌芽率低,萌芽开花延迟,花器官发育差,座果率低的问题。本试验以‘NU72’油桃为试材,观察了3种药剂对解除芽休眠的影响。结果表明,2％（NH2）2CS能提早花期,但存在药害现象,6％KNO3不能提早花期,并且花期不整齐,5％NH4NO3效果与6％KNO3类似。同时化学药剂处理促进花芽内H2O2的积累,抑制了过氧化氢酶（CAT）活性但促进了过氧化物酶（POD）活性,超氧物岐化酶（SOD）活性变化较小,化学药剂使花芽的呼吸速率增加,其中磷酸戊糖途径（PPP）代谢增加,糖酵解（EMP）降低,而三羧酸循环（TCA）代谢波动较小,葡萄糖－6－磷酸脱氢酶（G6PDH）活性在化学药剂处理时也增加。     

Comparative temporal analyses of the <Emphasis Type="Italic">Pinus sylvestris</Emphasis> L. var. <Emphasis Type="Italic">mongolica litv.</Emphasis> apical bud proteome from dormancy to growth

Bud dormancy in perennial plants adapts to environmental and seasonal changes. Bud dormancy is of ecological interest because it affects forest population growth characteristics and is of economical interest because it impacts wood production levels. To understand Pinus sylvestris L. var. mongolica litv. bud-dormancy and bud-burst mechanisms, we characterized the proteomes of their apical buds at the four critical stages that occur during the dormancy-to-growth transition. Ninety-six proteins with altered expression patterns were identified using NanoLC–ESI-MS/MS. The majority of these proteins (57%) are involved in metabolic and other cellular processes. For 28% of the proteins, a function could not be assigned. However, because their expression levels changed, they may be potential candidate bud development- or dormancy-related proteins. Of the 75 non-redundant bud proteins identified, ascorbate peroxidase, pathogenesis-related protein PR-10, and heat shock proteins dramatically increased during August and November, suggesting that they may involved in the initiation of bud dormancy. Conversely, S-adenosylmethionine synthetase, abscisic acid/stress-induced proteins, superoxide dismutase (SOD), caffeoyl-CoA O-methyltransferase, actin, and type IIIa membrane protein cp-wap13 had greater expression levels during April, suggesting that they may be involved in the initiation of bud dormancy-release. Cell division cycle protein 48 and eukaryotic initiation factors 4A-15 and 4A had greater expression levels during May, suggesting that they may regulate cell proliferate and differentiation in the shoot apical meristem. These observations provide insights into the molecular mechanisms that induce or break bud dormancy.     

Change in plasma membrane ATPase activity during dormancy release of vegetative peach-tree buds

Plant dormancy and dormancy breaking depend, at least partially, on peculiar short distance relationships between buds and tissues underlying buds (bud stands). In peach-tree, it was previously observed that dormancy was related to a high nutrient absorption capacity in tissues underlying buds. This situation could be linked to higher plasma membrane ATPase activity (EC 3.6.1.3), inducing a higher nutrient absorption, in bud stands. This work consists of characterization of the plasma membrane ATPase activity in vegetative buds and bud stands during the rest period and dormancy release. During the dormant period (October and November), plasma membrane ATPase activity was found to be higher in bud stands than in buds. This was correlated with a lower amount of plasma membrane ATPase in buds compared to bud stands during this period. Moreover, plasma membrane ATPase activation by trypsin treatment was not the same in both tissues and different levels of ATPase activation could be noted within the same tissue during the different stages of dormancy release. According to these results, it can be postulated that dormancy release in peach-tree, is related to modifications of plasma membrane ATPase properties in buds and bud stands during winter time.     

KNAP2, a class I KN1-like gene is a negative marker of bud growth potential in apple trees (Malus domestica [L.] Borkh.)

The determinism of bud bursting pattern along the 1-year-old shoot was studied at the molecular and morphological levels in the apple tree variety 'Lodi' which shows an acrotonic tendency. At the molecular level, the expression of KNAP2, which belongs to the class I KN1-like gene family, was studied. Measurements were carried out during dormancy (October), breaking dormancy (January) and just before bud bursting (March). The results showed that KNAP2 is more highly expressed in buds that will remain at rest in the spring. Expression of KNAP2 was found in the meristem and in the marginal meristem of the two latest shaped primordia. In the January and March buds, this gene is also expressed in the procambial zone underneath the apical meristem. This study therefore suggests that KNAP2 may be considered as a negative marker of bud growth potential and that the growth inhibition in proximal buds could partially result from differential gene activity. At the morphological level, it was shown that no organogenetic activity took place between October and March as revealed by the constant number of leaf primordia in buds. Nevertheless, those buds likely to grow the following spring had a larger size and fewer hard scales than other buds. This suggests that genetic control may act together with other mechanisms, possibly physical (number of scales) or biochemical, to control bud inhibition.