水稻叶片中活性甲基循环、转移相关基因对干旱胁迫的应答

 以中旱3号、汕优63和矮仔占为材料,在10% PEG6000模拟干旱条件下,分别应用基因芯片和mRNA 差异显示技术检测了不同基因型水稻叶片中活性甲基循环、转移相关基因表达对干旱胁迫的应答。中旱3号和汕优63叶片中活性甲基循环过程受干旱胁迫诱导,而矮仔占叶片中这一过程受到干旱胁迫的抑制;干旱胁迫诱导中旱3号叶片中更多甲基转移酶基因的表达,尤其是对Rubisco蛋白甲基化修饰基因转录的诱导,有利于防止Rubisco蛋白的氧化降解;干旱胁迫抑制矮仔占叶片中DNA甲基转移酶、组蛋白甲基转移酶基因的转录。这一结果证明水稻中甲基循环、转移基因的表达在水稻耐旱中有一定的作用。
     

Photoinhibitive and Recovery Properties of Hybrid RiceⅡ You 129 under Field Conditions

Photoinhibitive properties of super-high-yielding hybrid riceⅡ you 129 and its adaptation mechanism to strong light stress were investigated by measuring the light-response curve, diurnal variations of net photosynthetic rate and chlorophyll fluorescence parameters of Ⅱ you 129 leaves and compared with Shanyou 63. Photoinhibition of rice flag leaves under field conditions mainly resulted from the increase of thermal dissipation, especially for thermal dissipation depended on the xanthophyll circle, but no destruction of photosynthetic apparatus occurred. Potentially super-high-yielding hybrid rice Ⅱ you 129 was more tolerant to photoinhibition than Shanyou 63, because it had higher light saturation intensity and maximum net photosynthetic rate; more active xanthophyll cycle, and more rapid recovery ability after photoinhibition.     

自然条件下杂交水稻Ⅱ优129的光抑制及其恢复特性

通过测定田间高产杂交籼稻Ⅱ优129与对照汕优63的光响应曲线、光合速率和叶绿素荧光参数日变化,探讨了Ⅱ优129剑叶光合作用的光抑制特性及其对强光的适应机制。结果表明, 田间自然条件下杂交水稻叶片发生光抑制现象的主要原因是热耗散的增加,尤其是依赖叶黄素循环的热耗散的增加,而不涉及光合机构的破坏。具有超高产潜力的水稻品种Ⅱ优129比对照汕优63耐光抑制,其原因主要是Ⅱ优129具有较高的饱和光强、最大光合速率和叶黄素循环活力,遭受光抑制伤害后的恢复能力较强。     

根际促生细菌对干旱胁迫下水稻生理特性的影响

【目的】本研究旨在探究植物根际促生菌蜡状芽孢杆菌F06菌株对不同干旱胁迫下水稻汕优63生理特性的影响。【方法】在盆栽试验条件下,以水稻汕优63为种植材料,研究了轻度(LD)、中度(MD)、重度(SD)3个干旱强度下接种蜡样芽孢杆菌(Bacillus cereus)F06对水稻生理特征的影响。【结果】与正常水分管理相比,干旱胁迫(DS)下水稻叶片光合速率(P_n)和气孔导度(g_s)逐渐降低;而干旱胁迫下接种F06可显著减缓P_n和g_s下降,与不接种(NP)处理相比,P_n和g_s分别增加7.67%、12.97%、18.14%和11.51%、16.63%、17.07%,且呈现出随着干旱胁迫程度的提高,增幅增大的趋势。干旱胁迫下水稻叶片初始荧光(F_o)、非荧光淬灭系数(NPQ)显著上升,最大光化学效率(F_v/F_m)、光化学猝灭系数(q_P)显著下降;而干旱胁迫下接种F06可显著抑制F_o、NPQ升高和F_v/F_m、q_P降低,明显改善水稻叶片光能转换效率。干旱胁迫下接种F06虽然不能改变叶片水势、相对含水量和相对电导率的变化趋势,但可以有效降低其变幅。正常水分处理下接种F06虽然没有增加光合色素含量,但干旱环境下显著抑制了光合色素的分解或降低。干旱显著降低了水稻叶片和根系细胞分裂素(CTK)含量,增加了叶片中脱落酸(ABA)的含量;在干旱胁迫下,接种F06可显著提高叶片和根系中CTK的含量。【结论】由此可见,干旱生境下接种F06,可调节植物体内的激素含量,减少干旱胁迫下光合色素的分解或流失,提高光合速率,增强水稻在干旱环境中的适应能力。     

孕穗期干旱及复水对水稻叶片生理生化和产量的影响

现有关于水稻干旱胁迫期间各项生理生化指标的变化和对叶片细胞的伤害机理方面的研究已经比较丰富和全面,但是对于干旱胁迫结束复水后的生理变化研究却较少。本文以水稻品种中旱3号和黄华占为材料,在孕穗期进行淹水、轻度干旱、中度干旱和重度干旱4个处理,研究了干旱胁迫对水稻叶片生理生化性状（光合作用和抗氧化酶活性等）和产量的影响。结果表明,中度和重度干旱使水稻叶片光合速率下降12.9%~43.9%,这主要与气孔导度变化有关;处理结束复水后,中旱3号的生理生化指标均能恢复到正常水平,而黄华占在重度干旱后,其光合速率、气孔导度以及抗氧化酶活性不能再恢复;干旱处理导致水稻产量降低14.3%~47.9%,主要是由于有效穗数和结实率的降低。     

Proteomic Study for Responses to Cadmium Stress in Rice Seedlings

A proteomic approach including two-dimensional electrophoresis and mass spectrometric (MALDI-TOF MS) analyses was used to investigate the responses to cadmium (Cd) stress in seedlings of rice (Oryza sativa L.) varieties Shanyou 63 and Aizaizhan. Cd stress significantly inhibited root and shoot growth, and affected the global proteome in rice roots and leaves, which induced or upregulated the expression of corresponding proteins in rice roots and leaves when rice seedlings were exposed to 0.1 or 1.0 mmol/L Cd. The Cd-induced proteins are involved in chelation and compartmentation of Cd, elimination of active oxygen free radicals, detoxification of toxic substances, degradation of denatured proteins or inactivated enzymes, regulation of physiologic metabolism and induction of pathogenesis-related proteins. Comparing the Cd-induced proteins between the two varieties, the β-glucosidase and pathogenesis-related protein family 10 proteins were more drastically induced by Cd stress in roots and leaves of Aizaizhan, and the UDP-glucose protein transglucosylase and translational elongation factor Tu were induced by 0.1 mmol/L Cd stress in roots of Shanyou 63. This may be one of the important mechanisms for higher tolerance to Cd stress in Shanyou 63 than in Aizaizhan.     

土壤干旱胁迫对不同水稻品种叶片卷曲的影响

在人工控制土壤水分条件下,以13个具有不同耐旱性的栽培稻品种（组合）为供试材料,研究了在分蘖盛期土壤断水后随着干旱胁迫的加重叶片卷曲度（LRI）的动态变化及其与蒸腾、光合特性和叶片水分利用的关系。结果表明,不同品种的LRI明显增大的起始日期、明显增大期日均增量和最大值存在着显著差异,并依据叶片卷曲表现可将参试品种划分为敏感型（如中旱1号）、中间型（如旱稻8号）和钝感型（如巴西陆稻）三种类型。随着土壤断水后干旱胁迫的加重,LRI逐渐增大。LRI与气孔导度（SC）、蒸腾速率（TR）和净光合速率（Pn）的关系因品种类型而异;LRI每增加1级,中旱1号、旱稻8号和巴西陆稻的SC分别降低0.26、0.20和0.17 mm/s,TR分别降低0.86、0.66和0.49 mmol/(m2·s);Pn呈显著下降的叶片卷曲度中旱1号明显小于巴西陆稻。当叶片适度卷曲时,WUE随LRI值的增大而提高,中旱1号LRI的变化同时引起TR和Pn显著改变进而影响WUE,而旱稻8号和巴西陆稻LRI变化对WUE的影响主要系TR改变所致。     

营养生长期持续高温处理对玉米叶片转录组及生化指标的影响

以玉米杂交种郑单958和登海605为材料,通过转录组测序分析玉米营养生长期叶片受热胁迫后基因差异表达、基因功能富集和KEGG通路富集,使用微量法和HPLC法分析叶片中相关生化指标的变化。结果表明,高温胁迫会降低光合作用关键酶二磷酸核酮糖羧化酶(Rubisco)和三羧酸循环限速酶柠檬酸合酶(CS)的活力,升高丙二醛(MDA)、脱落酸(ABA)和赤霉素(GA₃)的含量,降低水杨酸(SA)的含量。与常温对照比,玉米营养生长期叶片受热胁迫后,两个品种4个比较组合累计检测到16 996个差异表达的基因,共同差异表达的基因有1 160个。光合作用、细胞代谢、叶绿体膜和质体膜等相关基因的表达会响应热胁迫。     

ABA对水分胁迫下水稻籽粒灌浆的调节

 利用水平分根系统研究了水稻抽穗后水分胁迫条件下ABA对水稻品种汕优63叶片光合特性、糖代谢和籽粒灌浆过程的调节。当一半根系受干旱胁迫时，叶片的水势和相对含量变化很小，但叶片和籽粒内ABA的含量显著增加。整个根系受干旱胁迫时，叶片的水势和相对含水量随着胁迫的增加而显著地降低，ABA含量则显著增加。半干和全干处理的水稻叶片的光合速率和RuBP羧化酶／氧化酶（Rubisco）的活性显著下降。叶片内蔗糖和淀粉的积累和代谢与叶片内ABA的含量和水分状况存在密切的关系，低浓度的ABA和高的叶水势与高的蔗糖积累和蔗糖磷酸合成酶活性相一致，但当叶片ABA的浓度增加时，即使叶片的水势与对照相似，叶片的蔗糖和淀粉积累以及蔗糖磷酸合成酶的活性则显著地下降，叶片的α－和β－淀粉酶活性似与ABA和水分状况关系不密切。ABA对籽粒灌浆有显著的抑制作用，ABA对籽粒灌浆的抑制作用至少部分与其对催化水稻籽粒内蔗糖代谢和淀粉积累的关键酶蔗糖合成酶的抑制有关。另外，干旱处理对叶片光合速率和蔗糖与淀粉积累的抑制程度显著高于半干旱处理，说明干旱本身也对这些过程有直接的效应。根据这些结果，作者认为，当水稻遇到干旱胁迫时，ABA对其光合作用、糖代谢以及籽粒的发育过程都有极其显著的调节作用。还就ABA对这些生理过程调节的可能性进行了讨论     

一个水稻NADPH氧化还原酶相似基因的克隆及表达分析

采用荧光差显(fluorescent differential display,FDD)技术,比较分析了干旱处理与未处理水稻叶片及根中的mRNA表达,并利用H. A. Yellow PAGE(含0.1% H. A. Yellow)再分离与大矩阵(macroarray)筛选相结合的方法,发现1个与拟南芥NADPH氧化还原酶高度相似(96%)的差异片段。该基因的cDNA全长为1423 bp,编码一个具有345个氨基酸残基的多肽。在正常情况下该基因表达量很低,而在干旱处理中高表达。讨论了干旱胁迫下NADPH氧化还原酶基因的可能功能。     

实时PCR定量分析干旱胁迫下水稻糖原合成酶激酶基因表达差异

通过实时荧光定量PCR进行相对定量分析,得到水稻IR64糖原合成酶激酶（glycogen synthase kinase, GSK）基因在不同生育期、不同组织干旱胁迫处理前后的表达差异。 叶片中GSK基因为组成型表达,各生育期干旱胁迫时诱导合成增加,尤其从分蘖期到抽穗期表达更为显著,说明这段生长期叶片对水分缺乏最敏感;苗期根系处理后GSK表达明显下降,叶片处理前后GSK表达无明显变化。由此可见,GSK基因的表达在不同的发育时期、不同的组织中呈现差异。     

转基因杂交稻组合汕优63和汕优559对白叶枯病的抗性

 采用包括与Xa21相匹配的鉴别菌系在内的12个国际鉴别小种和中国7个病原型，接种由明恢63、盐恢559的Xa21转基因单拷贝抗性纯合系与珍汕97A杂交获得的转基因杂交稻汕优63 (汕优63-Xa21 73个 F1群体和转基因杂交稻汕优559（汕优559-Xa21 13个F1群体，测定转基因杂交稻对白叶枯病的抗性。结果表明这2个转基因杂交稻组合具有优良的广谱抗性， 抗谱与Xa21基因供体IRBB21相同，非转基因杂交稻表现感病。     

Identification of Differentially Expressed Genes Induced by Ammonium Nitrogen in Rice Using mRNA Differential Display

RNAs isolated from ammonium- and nitrate-treated rice leaves were used to screen differentially expressed genes through mRNA differential display. A total of 72 bands appeared significant differences and some of them were further confirmed by reverse Northern and Northem blot. The results showed that two genes, A-02 (Oryza sativa drought stress related mRNA) and A-03 (Zea mays partial mRNA for TFIIB-related protein) were highly up-regulated in the ammonium-fed rice leaves. The enzyme assays showed that the activities of the two anti-oxidative enzymes, catalase and peroxidase, and the content of a non-enzymic antioxidant, glutathione, were significantly higher in the ammonium-fed rice leaves than those in the nitrate-fed ones, indicating that the ammonium nutrition might be beneficial for rice plants to improve the stress resistance during growth and development.     

孕穗期水稻不同功能叶的发育蛋白质组学分析

 从蛋白质组学角度对杂交水稻汕优63孕穗期4个叶位功能叶片蛋白质组变化进行了研究,以揭示水稻孕穗期叶片蛋白质组的表达差异。4个不同时期功能叶片蛋白质经双向电泳得到分离,应用Imagemaster 2D Elite 5.0软件对所得到的双向电泳图谱进行分析比较,共得到差异表达蛋白质点23个,差异表达蛋白质点经质谱分析,有14个得到鉴定。对鉴定出的蛋白功能进行研究,结果表明:差异表达的蛋白质大部分(9个)为参与光合作用和呼吸作用的蛋白质;其次为与蛋白结构及功能调控相关的蛋白质(3个),表达量表现为随叶位上升而上调;其余为参与氨基酸代谢的蛋白质(2个),表达量在较低叶位叶片中无显著规律,在剑叶中的表达量显著上调。蛋白质是细胞功能执行者,受细胞功能或细胞环境变化的影响,因此,上述蛋白质发生变化的原因应与水稻不同叶位叶片生长发育特性有关。     

低温对杂交水稻及其亲本灌浆期剑叶果糖1，6-二磷酸酯酶活性的影响

Effects on chilling on fructose 1,6-diphosphatase (FBPase) of flag leaves in five hybrids Shanyou 63, Shanyou 64, Weiyou 64, Qingyouzao and Xiuyou 57 were investigated. The results indicated that chloroplast FBPase activity of flag leaves in hybrid rice F₁ and their parents at filling stage decreased after low temperature (1℃) treatment. The sensitivity of FBPase to chilling can reflect difference of cold tolerance among these hybrids. The order of cold tolerance was as follows: Japanica type Xiuyou 57, Indica type Qingyouzao, Shanyou 63, Shanyou 64. and Weiyou 64. The cold tolerance of F₁hybrid rice was inherited toward to that of the maternal line. The possible role of FBPase during grain filling and the relationship between FBPase activity of flag leaves and cold tolerance in hybrid rice were discussed.     

茶树钙依赖性蛋白激酶基因CsCDPK17的鉴定及表达分析

钙依赖型蛋白激酶(Calcium-dependent protein kinases,CDPKs或CPKs)是植物细胞中重要的钙离子信号受体,普遍参与了植物生长发育和胁迫响应等调控过程。本研究从茶树龙井43中克隆到1条CDPK基因,经测序验证该序列包含1611bp的完整ORF,编码536个氨基酸。结合序列比对和进化分析发现该蛋白N-末端具有豆蔻酰化位点,具备钙依赖性蛋白激酶的保守结构域,并且与拟南芥AtCDPK17的同源关系最近,因此命名为CsCDPK17(Genbank登录号为:MK238482)。蛋白质理化特性分析发现其蛋白分子量为59.9kD,理论等电点pI为5.43,属无跨膜结构域的亲水性蛋白。使用水稻原生质体和烟草叶片两种瞬时表达的方法均证明CsCDPK17是细胞质膜和细胞核双定位蛋白。对克隆到的CsCDPK17上游2000bp的启动子区分析发现了多个与基因转录、光、激素(ABA、SA、MeJA等)相关的顺式作用元件。表达分析显示,CsCDPK17在成熟叶和种子中表达水平较高,而在根中的表达水平最低;在龙井43、大面白等4个品种冷驯化过程中,该基因的表达随着冷驯化过程显著上调,随着脱驯化过程下调;同时还发现CsCDPK17能够不同程度地响应低温(最高5.1倍)、干旱(最高2.3倍)、渗透(最高2.4倍)胁迫。本研究的结果表明CsCDPK17在茶树的生长发育和低温、干旱、渗透等非生物胁迫响应的过程中均发挥一定的调控作用。     

Effect of Rolled Leaf Gene Rl（t） on Grain Quality in Hybrid Rice

Effects of rolled leaf gene Rl（t） on grain quality characters of hybrid rice were analyzed by using three pairs of rolled leaf near-isogenic lines under two fertilizer treatments. Under normal fertilizer level （e.g. 450 kg urea per ha）, head rice rates and milled rice recovery of rolled leaf hybrids were significantly higher than those of corresponding non-rolled crosses, while the chalky rice rate and chalkiness were all lower. Of the RVA profiles, the peak viscosity, the hot paste viscosity and the breakdown viscosity of the rolled were all higher than those of the corresponding non-rolled ones to various degrees. Increasing fertilizer application for promoting panicle development increased the brown, milled and head rice rates except for Shanyou 63, furthermore, significant difference of head rice rates existed between the rolled leaf Shanyou 559 and Shanyou 559; while the peak viscosity, the hot paste viscosity and the breakdown viscosity all decreased to different levels; changes of values of other characters had no apparent regularity. It suggested that Rl（t） could improve rice quality under certain conditions.     

Simulation on Distribution of Photosynthetically Active Radiation in Canopy and Optimum Leaf Rolling Index in Rice with Rolling Leaves

By replacing leaf area index (LAI) with effective leaf area index (ELAI) through introduction of leaf rolling index (LRI), the distributions of photosynthetically active radiation (PAR) in the canopies of three hybrid rice combinations, Liangyou E32 with high LRI, Liangyoupeijiu with moderate LRI and Shanyou 63 with non-rolling leaves (normal), were simulated. The model based on ELAI could predict more accurately than that based on LAI. The PAR interception, conversion and utilization efficiency in the three combinations were studied to evaluate their optimal LRI and LAI. The PAR utilization efficiency of Liangyou E32 was lower due to excessive rolling leaves and less ELAI, and that of Shanyou 63 was also lower because of the faulty PAR interception and lower photosynthetic rate and saturation point at lower layer in canopy. Compared with the above two combinations, Liangyoupeijiu showed more appropriate distribution of PAR interception and conversion efficiency in canopy, and higher PAR utilization efficiency. The optimal LRI and LAI for Liangyoupeijiu were 0.11 and 7.6, respectively, which were close to the observed value, 0.11 and 7.9, respectively. However, the optimum LAI was 9.8 for Liangyou E32 and 6.2 for Shanyou 63, larger or smaller than those under the current plant density, which led to lower efficiency of PAR utilization. Besides, the optimum LRI for Liangyou E32 and Shanyou 63 were 0.12 and 0.08, respectively, which were close to the actual LRI for Liangyoupeijiu (0.11).     

甘蓝型油菜SERK基因家族的鉴定与系统进化分析

体细胞胚胎发生类受体激酶（somatic embryogenesis receptor-like kinase，SERK）与植物体细胞胚发生，尤其是与植物非生物逆境胁迫响应有关。为鉴定甘蓝型油菜中BnaSERK基因家族成员、揭示其进化关系及其与油菜盐/旱胁迫的响应，利用生物信息学方法对甘蓝型油菜品种中双11的SERK家族成员、基因结构、进化关系、选择压力等进行系统分析，并初步分析了部分BnaSERK基因在盐/旱胁迫下的表达响应模式。结果表明：在甘蓝型油菜基因组共鉴定到24个BnaSERK基因，它们不均等地分布在15条染色体上，可分为3个亚族，具有相对保守的基因结构和保守基序，且含多种与激素和非生物胁迫响应相关的顺式作用元件。共线性分析表明，甘蓝型油菜和拟南芥、白菜、甘蓝分别有14对、44对和32对基因表现出共线性。基于比较基因组学研究，甘蓝型油菜BnaSERK基因在经历多倍体化之后，出现了不同程度的丢失现象。分析表明，BnaSERK基因家族在芸薹属物种间的进化相对保守。盐/旱胁迫下的5个BnaSERK基因在叶片中的表达模式分析结果显示，BnaA07g29610D、BnaC01g43240D、BnaCnng07810D基因在盐胁迫下表达量上调，而BnaA01g23070D、BnaA07g23390D基因则表现为下调；BnaA07g23390D、BnaA07g29610D基因在干旱胁迫下表现为上调表达，而BnaC01g43240D、BnaCnng07810D、BnaA01g23070D基因呈下调趋势。表明BnaSERK基因可能在油菜响应盐/旱胁迫的调控机制中发挥着重要作用。