低温对不同基因型黄瓜叶片蛋白质组的影响

 【目的】分离不同基因型黄瓜叶片的低温诱导蛋白,为从蛋白质组角度揭示黄瓜应答低温逆境的调控机制打下基础。【方法】以黄瓜（Cucumis sativus L.）的日光温室品种‘新泰密刺’和露地栽培品种‘津研4号’为试材,采用分步提取法溶解叶片总蛋白质,运用SDS聚丙烯酰胺凝胶电泳、双向电泳、质谱分析与检索技术,在100 μmol?m-2?s-1的光照条件下,比较研究低温（15/15℃）和常温（25/18℃）下叶片蛋白质组的差异。【结果】利用提取液Ⅰ、Ⅱ和Ⅲ分步提取叶片总蛋白质后,提取液Ⅰ溶解的黄瓜蛋白质较少。对提取液Ⅱ和Ⅲ溶解的叶片蛋白质进行双向电泳后发现,相对于常温处理,低温使‘新泰密刺’的7个蛋白质点特异表达、2个蛋白质点上调表达、2个蛋白质点下调表达、5个蛋白质点未表达;使‘津研4号’的7个蛋白质点特异表达、1个蛋白质点上调表达、4个蛋白质点下调表达、9个蛋白质点未表达。低温下,‘新泰密刺’的蛋白质点3和4 以及‘津研4号’中蛋白质点2的表达量显著高于常温处理,通过MALDI-TOF质谱分析,这3个蛋白质点被鉴定为1,5-二磷酸核酮糖羧化酶的大亚基。【结论】黄瓜叶片的高亲水性蛋白质含量低;在相同的光照（100 μmol?m-2?s-1）条件下低温对不同基因型黄瓜叶片的蛋白质组均有影响;1,5-二磷酸核酮糖羧化酶可能与黄瓜叶片耐低温有关。     

黄瓜幼苗Rubisco与Rubisco活化酶对光强的响应

以津优3号幼苗为试材,研究弱光(LL:100 μmol· m-2·s-1)、亚弱光(SL:300 μmol·m-2·s-1)下黄瓜幼苗光合速率(Pn)、核酮糖-1,5-二磷酸羧化/加氧酶(Rubisco)、Rubisco活化酶(RCA)活性及其基因表达量的变化.结果表明,11d弱光处理后,黄瓜幼苗叶片的Pn、Rubis...     

Contamination of internationally traded wheat by herbicide‐resistant Lolium rigidum

As herbicide‐resistant weeds have spread in the agricultural fields of grain‐exporting countries, their seeds could be introduced into other countries as contaminants in imported grain. The spread of resistance genes through seed and pollen can cause significant economic loss. In order to assess the extent of the problem, we investigated the contamination by herbicide‐resistant annual ryegrass (Lolium rigidum) of wheat imported from Western Australia into Japan. Annual ryegrass seeds were recovered from wheat shipments and seed bioassays were conducted to identify resistance to the herbicides that are commonly used in Australia: diclofop‐methyl, sethoxydim, chlorsulfuron, and glyphosate. Nearly 4500 ryegrass seeds were detected in 20 kg of wheat that was imported in both 2006 and 2007. About 35% and 15% of the seeds were resistant to diclofop‐methyl, 5% and 6% were resistant to sethoxydim, and 56% and 60% were resistant to chlorsulfuron in 2006 and 2007, respectively. None was resistant to glyphosate in either year. As the contamination of crops by herbicide‐resistant weeds is probably a common phenomenon, the monitoring of incoming grain shipments is necessary to stem the further spread of herbicide‐resistant weeds into importing countries.     

不同施肥水平对南瓜RuBP羧化酶活性的影响

采用L^9（3^4）正交试验设计,系统的研究了不同施肥条件下南瓜穴盘苗的RuBP羧化酶活性。结果表明,氮素对南瓜穴盘苗的RuBP羧化酶活性影响最为显著。施用氮肥后,RuBP羧化酶活性显著降低;磷钾对南瓜穴盘苗的RuBP羧化酶活性影响最小,磷钾施用后,RuBP羧化酶活性与对照差异不显著。微肥对南瓜穴盘苗RuBP羧化酶活性的影响介于前两者之间,也使RuBP羧化酶活性较明显地降低。     

适度水分胁迫提高黄瓜幼苗光合作用弱光适应性

以黄瓜（Cucumis sativus L.）‘戴多星’品种为试材,在人工气候室内研究弱光（75 ~ 85 μmol · m-2 · s-1）下水分胁迫（40%基质最大持水量）对幼苗叶片光合、荧光特性及Rubisco含量的影响。结果表明,弱光逆境下,经20 d水分胁迫后的幼苗较正常水分条件下叶片光合色素（以单位面积计）含量提高,叶绿素a/b（Chl.a/b）下降,植株光饱和光合速率Asat、表观量子效率（AQY）、羧化效率（CE）、RuBP最大再生速率及光饱和点（LSP）均显著增加,CO2补偿点（CCP）显著下降,并且光系统Ⅱ（PSⅡ）最大光化学效率（Fv/Fm）、潜在光化学效率（Fv/Fo）、PSⅡ光化学量子效率（ΦPSⅡ）、光化学反应速率（Prate）、非环式电子传递速率（J）以及PSⅡ吸收光能向光化学反应分配比例P等叶绿素荧光参数呈增加趋势,PSⅠ和PSⅡ间激发能分配不平衡性（β / α–1）减小,叶片可溶性蛋白质含量和Rubisco大、小亚基含量上升,表明适度的水分胁迫有利于提高弱光下黄瓜幼苗的光合适应性;而正常光照下（500 ~ 600 μmol · m-2 · s-1）经水分胁迫后的黄瓜幼苗上述主要光合特性指标如Asat、RuBP最大再生速率、CE、ΦPSⅡ、Prate、J和P等有所下降,光系统间激发能分配不平衡性（β / α–1）增加,光合作用受到一定抑制。     

内源水杨酸参与黄瓜叶片光合系统对低温胁迫的响应

 为了探讨内源水杨酸（salicylic acid,SA）在黄瓜幼苗光合系统响应低温胁迫中的作用机制, 采用高效液相色谱法测定低温下黄瓜叶片中内源SA 含量的变化;通过SA 合成抑制剂Paclobutrazol（Pac, 100 μmol · L^-1）喷施和外源SA（50 μmol · L^-1）饲喂的方法调节内源SA 含量,并测定不同处理幼苗的叶 绿素荧光参数和光合碳同化关键酶基因的转录水平。结果显示：低温引起黄瓜幼苗内源SA 含量升高,Pac 预处理抑制SA 的积累。低温导致PSⅡ的最大光化学效率（F v/F m）、实际光化学效率（Φ PSII）、潜在光化 学活性（F v/F o）和光合电子传递效率（ETR）等降低,叶片光化学猝灭参数[（Y（NO）]升高;内源SA 含量降低使PSⅡ活性下降幅度增大,加重了叶片的光损伤程度。低温下PSⅡ吸收的光能分配于光反应的 部分减少,而以非光化学反应的过剩能量耗散Ex 为主要的光能分配途径,内源SA 含量降低会加剧光能 向Ex 的分配。低温时喷施Pac 的幼苗中Rubisco 小亚基基因（RbcS）和碳酸酐酶基因（CA）的表达水平 显著低于对照植株。对喷施Pac 的幼苗外源饲喂SA 后,内源SA 含量升高,低温下叶片光合活性得到有 效恢复,光损伤降低,光能分配趋于合理,RbcS 和CA 的表达水平升高。上述结果表明,低温下内源SA 的积累有助于维持黄瓜叶片中较高的光系统活性和碳同化能力,从而保护光合系统,降低低温胁迫对植 物的损伤。     

Magnesium deficiency–induced impairment of photosynthesis in leaves of fruiting Citrus reticulata trees accompanied by up‐regulation of antioxidant metabolism to avoid photo‐oxidative damage

Limited data are available on the physiological responses of leaves from fruiting trees to magnesium (Mg) deficiency. Magnesium deficiency–induced effects on photosystem II (PSII) photochemistry in leaves of fruiting (Citrus reticulate cv. Ponkan) trees were assessed by the chlorophyll a fluorescence (OJIP) transient. Magnesium deficiency decreased leaf CO₂ assimilation and carbohydrates, but had no effect on intercellular CO₂ concentration. Activity of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) and concentrations of Chlorophyll (Chl) and carotenoids (Car) decreased to a lesser extent than CO₂ assimilation. Chlorophyll a fluorescence transient from Mg‐deficient leaves had increased O step and decreased P step, accompanied by positive ΔL, ΔK, ΔJ, and ΔI bands. Magnesium deficiency decreased maximum quantum yield of primary photochemistry (F_v/F_m), quantum yield of electron transport from Q<$>_A^‐<$> to the photosystem I (PSI) end electron acceptors (φ_R0), maximum amplitude of IP phase and total performance index (PI_{tot, abs}), but increased deactiviation of oxygen‐evolving complex (OEC) and energy dissipation. Magnesium‐deficient leaves had higher or similar activities of antioxidant enzymes except for lower catalase (CAT) activity, higher or similar concentrations of antioxidant metabolites, and a higher ratio of Car : Chl. Magnesium‐deficiency did not affect concentration of malondialdehyde (MDA) and ratios of ascorbate (ASC) to ASC + dehydroascorbate (DHA) and reduced glutathione (GSH) to GSH + oxidized glutathione (GSSG). In conclusion, Mg deficiency–induced impairment of the whole photosynthetic electron transport chain may be the main factor contributing to decreased CO₂ assimilation. Enhanced energy dissipation and antioxidant metabolism provide sufficient protection to Mg‐deficient leaves against photo‐oxidative damage.     

胡麻异质型ACCase亚基基因的克隆与表达分析

为研究胡麻异质型乙酰辅酶A羧化酶BCCP、BC、α-CT和β-CT四个亚基基因的生物学功能,采用基因克隆、生物信息学和RT-PCR技术分别对胡麻accA、accB、accC和accD 4个基因进行分析。结果发现陇亚10号accA基因编码α-CT亚基,CDS序列全长为2364 bp,编码787个氨基酸;accB基因编码BCCP亚基,CDS序列全长为1173 bp,编码275个氨基酸;accC基因编码BC亚基,CDS序列全长为1574 bp,编码527个氨基酸;accD基因编码β-CT亚基,CDS序列全长为1137 bp,编码378个氨基酸,且4个基因编码的蛋白都是脂肪酸系数较高的亲水性蛋白。RT-PCR结果表明,胡麻accA、accB、accC和accD 4个基因在3个胡麻品种(高含油量的张亚2号、中含油量的陇亚10号和低含油量的R2-17)的不同时期不同组织的表达模式不同,accA、accB、accC和accD四个基因在R2-17、陇亚10号和张亚2号3个胡麻品种所有时期的不同组织中均有表达,但在种子中的表达量均明显高于其他组织,尤其在种子发育10~25 d油脂快速积累时期,4个基因的表达量都迅速增加并达到最高峰。研究表明异质型乙酰辅酶A羧化酶基因可能调控胡麻种子发育前期油脂的合成积累。     

木兰门作物RubisCO大亚基的分子适应性进化研究

分子进化中发生正选择的一个标志是密码子的错义替换（dN,氨基酸替代）比同义替换（dS,氨基酸不变）有更高的替换速率,计算dN与dS之比ω能衡量蛋白质的进化趋势和所受到进化压力的大小。ω=1、〈1和〉1分别说明蛋白质在进化过程中受到中性选择、负选择和正选择。