NO对生长发育中棉花叶片NO含量及其对抗氧化物酶的影响

以早衰性状不同的棉花栽培品种为材料,在自然条件下和外施一氧化氮(nitric oxide, NO)的条件下,调查早熟棉花植株真叶和子叶衰老过程中NO含量变化和抗氧化酶活性及相关基因的表达。结果表明,大田条件下,NO含量在幼嫩叶片中最高,随着叶片的衰老含量逐渐降低;在叶片发育后期早衰材料的NO含量下降快,并且显著低于不早衰材料。室内条件下,植株发育过程中,NO含量在幼嫩子叶中最高在生长后期最低;外施SNP溶液后的植株,其NO含量在子叶的整个生育期都比对照组高,且两者差异显著。对照组和处理组的过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)的活性及相关基因的表达第7天较低,第14天最高,随后逐渐下降;在同一时期,处理组显著高于对照组,在生育后期表现的更为明显。外施SNP可显著降低参试品种过氧化物酶(POD)的活性和相关基因的表达。在子叶发育初期,外源NO对超氧化物歧化酶(SOD)的活性有抑制作用,随着叶片衰老,处理组的SOD活性又高于对照组。不同类型的SOD对NO的反应不同,Cu/Zn SOD最敏感,其中又以cCu/Zn SOD基因的作用更突出。NO通过调控植株体内CAT、APX、POD和SOD等氧化/抗氧化系统,延缓叶片的衰老进程。     

陆地棉小GTP结合蛋白基因GhRop4的克隆及其表达分析

【目的】对陆地棉小GTP结合蛋白基因(Small GTP-binding protein)GhRop4(AY965614.1)在多种逆境胁迫下的应答表达模式进行研究分析,为棉花抗逆相关基因克隆及棉花抗逆分子机制研究奠定基础。【方法】利用生物信息学方法分析了GhRop4基因的结构特征和进化树关系,并通过荧光定量PCR(Real-time quantitative polymerase chain reaction,qRT-PCR)的方法分析Gh Rop4基因的组织表达特异性和不同逆境诱导条件下的表达模式。【结果】以陆地棉c DNA为模板克隆得到GhRop4基因,其开放阅读框为588 bp,编码包含195个氨基酸残基的Ⅰ型Rop蛋白。氨基酸多重序列比对表明,GhRop4与其它植物Rop蛋白高度同源,符合Rop蛋白结构特点。qRT-PCR分析表明,GhRop4基因在棉花幼苗根、茎、叶、子叶和下胚轴中均有表达,且在子叶和茎中表达水平较高。GhRop4基因对高盐、干旱、低温和棉花黄萎病菌处理都有一定程度的响应。【结论】Gh Rop4基因在陆地棉的逆境胁迫适应过程中可能具有重要的作用。     

一个水稻叶片衰老上调表达基因的初步生物学功能分析

叶片衰老是其发育的最后阶段。通过对水稻叶片衰老机制的研究, 有计划地控制或延缓衰老的发生具有重要的理论价值和实践意义。本研究基于基因表达芯片数据挑选了一个叶片衰老上调表达候选基因A12 (LOC_Os 07g41230)。A12基因在水稻全生育期表达谱数据库中的表达模式及在水稻抽穗后不同时期剑叶中的表达量检测结果进一步证明其确为叶片衰老上调表达基因。生物信息学预测A12基因启动子区域存在大量的与激素诱导相关的顺式作用元件。实时定量PCR结果显示A12基因对茉莉酸(JA)和激动素(KT)的诱导有明显的响应, 但是对油菜素内酯(BR)、赤霉素(GA)、生长素(IAA)及脱落酸(ABA)的诱导则无明显响应。对A12基因对应的水稻T-DNA插入突变体观察发现, A12基因的突变会导致剑叶早衰。这些结果为进一步深入研究A12基因的生物学功能打下了基础。     

小麦脱植基相关基因在春性小麦叶片叶绿素降解过程中的作用分析

小麦叶片的衰老会导致产量的损失,而叶绿素降解是小麦叶片衰老的明显特征,分析小麦叶绿素降解过程中脱植基反应的相关基因叶绿素酶（TaCLH）和脱镁叶绿素水解酶（TaPPH）在春性小麦叶片衰老过程中的作用,为解析小麦叶绿素降解的分子机制提供参考。以10个春性小麦品种为参试材料,对衰老过程中TaCLH和TaPPH的相对表达量进行测定,结合不同品种在开花后不同时期的相对叶绿素含量（SPAD）、功能绿叶面积（GLAD）和叶绿素荧光参数的变化规律,研究叶片衰老过程中TaCLH和TaPPH与SPAD、GLAD和叶绿素荧光参数的相关关系。结果表明,TaPPH的相对表达量与GLAD、SPAD及叶绿素荧光参数[ETR、F_v/F_m、Y(Ⅱ)]等生理指标之间存在极显著负相关关系,与TaCLH相对表达量存在极显著正相关关系,表明TaPPH在春小麦叶绿素降解过程的脱植基反应中起主要作用。     

Morphogenesis of Pigment Glands and Metabolic Characteristics of Gossypol in Gossypium bickii

[Objective]We studied the process of pigment gland formation and the dynamic change of gossypol synthesis, the dynamic relationship and molecular mechanism between the pigment glands and gossypol during the process of embryo formation and seed germination in G. bickii Prokh. [Method] We chose G. bickii as meterial and studied the tissue section observation, determination of gossypol contents and gene expression profile analysis of pigment gland formation and gossypol synthesis genes. [Result] The results showed that a large number of pigment protoglands formed on the cotyledons during embryo development process in G. bickii. The pigment protoglands formed to cavity structure at 12 hours after seed germination, and then converted into mature pigment glands. Analysis of gene expression profiles related to the pigment gland formation and gossypol synthesis during seed germination showing that GoPGF gene was highly expressed on the cotyledons of seed germination of 12 hours, which may be related to pigment protoglands forming to cavity structure of G.bickii. Farnesyl pyrophosphate synthase gene (FPS) was highly expressed on the cotyledons of seed germination of 24 hours showing it may be associated with the gossypol transported into the cotyledon pigment glands. CYP706B1 gene was highly expressed on the cotyledons of seed germination of 0 hours and HMG1 were highly expressed on the cotyledons of seed germination of 0 hours, 36 hours and 48 hours showing these genes may be associated with the formation of pigmented cavities and the storage and transport of gossypol; Cadinene synthase genes(CAD1-A) and HMG2 genes were not closely related to the pigment gland formation. [Conclusion] The 12 hours after seed germination is the key period of the morphogenesis of the pigment glands, and the morphogenesis of the pigment glands is synchrony with the time of the occurrence of gossypol in G.bickii. GoPGF, FPS, CYP706B1 and HMG1 are associated with the formation of pigmented cavities and the storage and transport of gossypol.     

陆地棉转录因子GhNAC78基因的特征及功能分析

本实验室从陆地棉中克隆得到GhNAC78,分析了其基因特征及功能。该基因cDNA全长为937 bp,开放阅读框为876bp,编码291个氨基酸,隶属于NAM转录因子亚家族。体外转录激活实验表明,GhNAC78具有转录激活活性。上游启动子区1537 bp序列的生物信息学分析可知其包含多种激素、胁迫和光响应元件,表明GhNAC78广泛地参与植物生长发育和胁迫响应的调控。荧光定量结果显示,GhNAC78在棉花叶片衰老过程中高调表达,推测其参与叶片衰老的调控。     

粳稻根系与叶片早衰的关系及其对籽粒灌浆的影响

以衰老程度不同的品种为试材,对根系和叶片衰老的生理机制、根系与叶片衰老的关系、根系和叶片衰老对籽粒灌浆结实的影响进行了研究.结果表明:(1)抽穗后根系和叶片MDA含量持续上升,而SOD活性却先上升然后再持续下降,说明MDA含量上升最初并不是由SOD活性下降引起的.(2)根系和叶片衰老受ZRs和ABA平衡关系的调控,ZRs含量的下降和     

小麦旗叶衰老过程不同数学模型拟合比较及衰老特征分析

为了准确了解小麦叶片的衰老特征,筛选适合描述小麦叶片衰老过程的数学模型, 2011年和2012年分别以91个和105个小麦品种(系)为材料,用"S"型曲线中的Logistic、Gompertz和Richards模型拟合了试验品种叶片的衰老过程,解析了其叶片衰老特征。结果表明,试验品种可分为延绿、中等延绿、中等早衰和早衰4种类型。其旗叶衰老过程可分为衰老起始期、快速衰老期和衰老结束期3个阶段,3个阶段旗叶的衰老速度表现为"慢–快–慢",不同延绿类型品种开花后旗叶的绿色叶面积百分比下降主要在衰老过程的中后期。3种模型对不同延绿类型品种旗叶衰老过程均可以拟合,Gompertz和Richards模型拟合度接近,高于Logistic模型。Gompertz模型的拟合度以早衰中等早衰中等延绿延绿类型。不同延绿类型品种旗叶衰老曲线特征参数达到最大衰老速度时间(TMRS)、平均衰老速度(ARS)和绿色叶面积持续期(GLAD)存在显著差异, TMRS和GLAD以延绿中等延绿中等早衰早衰, ARS以早衰中等早衰中等延绿延绿。Gompertz模型对小麦叶片衰老过程的拟合度优于Logistic模型。叶片衰老过程特征参数可以用于品种延绿性差异评价。     

Responses and Underlying Mechanisms of Different Mainstem Leaves on Cotton to Defoliant Thidiazuron

[Objective] This study elucidates the responses of different mainstem leaves on cotton to defoliant thidiazuron and reveal the underlying mechanisms. [Method] Cotton plants were raised hydroponically. At the ten-leaf stage, 4.54 mmol·L^－1 thidiazuron was evenly applied to all mainstem leaves, the youngest full-expanded leaf (the fourth leaf from apex) and the youngest leaf (the first leaf from apex). Then, the progress of abscission zone formation and shedding of each leaf was observed, and the expression level of genes related to ethylene synthesis and signal transduction in the fourth and first leaf from apex was analysed. [Result] The abscission zone developed in the following sequence: the first leaf from the top, then the lowest leaves (the ninth or tenth leaf from the top, 35-40 days old), and the fourth leaf from the top. However, the leaf abscission was in a different order: the earliest was the first leaf from apex, then the fourth leaf from apex, and the latest was the lowest leaves which were prone to wither and stick to the stalk. The GhACS1 expression was up regulated in both the first leaf and the fourth leaf from apex within 24 h after thidiazuron treatment. However, the expression of this gene changed more rapidly and sharply in the first leaf from the top than that in the fourth leaf from the top. In addition, the expression of GhACO1 (ethylene synthesis gene), GhEIN3 and GhERF23 (ethylene signal transduction genes) was significantly increased up to ten times in the first leaf from the top, whereas the expression of these genes did not change in the fourth leaf from the apex. [Conclusion] After exposed to defoliant thidiazuron, the time of the abscission zone formation of different leaves on cotton main stem is different, and the order of shedding is not consistent with that of abscission zone formation. The youngest ready to expand leaf abscises earlier than the youngest fully expanded leaf after thidiazuron treatment, which is possibly associated with the rapid expression change of genes related to ethylene synthesis and signal transduction in the former.     

陆地棉GhERF8基因的克隆与表达分析

 乙烯响应元件结合因子（Ethylene-responsive element-binding factor,ERF）是植物中最大的转录因子家族之一。本研究以棉花叶片cDNA文库为基础,从陆地棉中棉所10号中克隆得到一个新的ERF基因,命名为GhERF8（GenBank：JN656957）。该基因编码265个氨基酸,蛋白序列中包含一个AP2保守结构域。采用荧光定量PCR（RT-PCR）的方法,对GhERF8基因在棉株不同生育时期叶片和不同组织中的表达水平进行了定量分析。结果表明,GhERF8基因在各组织中均有表达,但在成熟后期的叶片中表达量最高。GhERF8表达量与叶片衰老过程中叶绿素含量出现相反的变化趋势,推测GhERF8可能与叶片衰老有一定关系。在乙烯利和茉莉酸处理下GhERF8基因在叶片中上调表达,而在脱落酸处理下GhERF8表达量无显著变化,推测GhERF8可能处于乙烯和茉莉酸信号转导网络中,且表达途径为非ABA依赖途径。     

山核桃水溶性成分中神经营养活性因子的分离与鉴定

本研究旨在揭示山核桃水溶性成分的神经营养活性及活性因子。利用脱脂山核桃水溶性提取物处理SH-SY5Y细胞,用倒置相差显微镜观察细胞突触生长,用qPCR检测相关基因表达,使用HPLC 及LC-MS对山核桃水溶性成分神经营养活性因子进行分离鉴定。结果显示,山核桃的水溶性成分能够显著促进SH-SY5Y细胞的突触生长,并可以显著上调SH-SY5Y细胞NF160 基因的表达。HPLC及LCMS检测结果发现其水溶性成分主要组成为胞苷、尿苷、鸟苷、腺嘌呤、腺苷等。由此可知,山核桃水溶性成分通过上调NF160基因表达具有神经营养活性。     

棉花子叶衰老过程中氮素的再利用

 以陆地棉早熟品种中棉所58和中熟品种邯郸109为材料,研究子叶衰老过程中氮素的再利用率（NRE）及氮代谢相关指标的变化。结果表明：子叶衰老过程中,单位面积总N含量（NPA）逐渐降低,中棉所58 NRE为69.26%,邯郸109为54.45%;叶绿素、可溶性蛋白质含量和硝酸还原酶（NR）活性先上升后降低,NR最先降低,中棉所58和邯郸109分别在9 d、14 d开始下降;中棉所58各指标比邯郸109早下降4～8 d。谷氨酸脱氢酶（GDH）活性先下降后上升,表明GDH可能是氮素再利用的关键酶。     

Cytokinins: A key player in determining differences in patterns of canopy senescence in Stay-Green and Fast Dry-Down sunflower (Helianthus annuus L.) hybrids

Leaf senescence during grain filling can reduce crop yield. We studied, under field conditions and during grain-filling, the association between leaf cytokinin levels and the onset of leaf senescence in sunflower hybrids of contrasting canopy senescence patterns (Paraiso75, stay-green [SG] and Paraiso65, fast dry down [FDD]). At crop level, dynamics of live root length density (LRLD) and green leaf area index (GLAI) were followed, while at leaf level dynamics of total chlorophyll content, trans-Zeatin content, net photosynthesis and PSII quantum yield, were followed in leaf positions 17, 20, 22 and 24. Responses of these leaf variables to exogenous cytokinin applications to leaves at position 17 were also followed. SG exhibited greater (p < 0.05) LRLD and GLAI values at anthesis. In both hybrids, LRLD began to fall before GLAI. All variables decreased earlier (p < 0.05) in FDD. Initial leaf levels of trans-Zeatin were three times higher (p < 0.05) in SG. Exogenous cytokinin applications maintained leaf-level variables. These are the first results showing associations between LRLD dynamics with the dynamics of leaf cytokinin levels and changes in indicators of leaf functionality. Also, this is the first study in which estimates are made of cytokinin thresholds below which leaf senescence begins in two hybrids of contrasting canopy senescence patterns. These advances in the understanding, at both crop and leaf levels, of the controls and consequences of SG during grain filling, a trait known to improve crop water uptake under drought and increase biomass accumulation during grain filling, provide support for breeding efforts aimed at profiting from this trait to increase crop yields.     

棉花不同叶位主茎叶衰老特征的研究

 在大田条件下,研究了棉花不同叶位主茎叶在盛铃末期的衰老状况。以转基因抗虫棉33B为材料,测定全部主茎叶的主要生理生化指标,以分析不同部位叶片衰老的差异。结果表明,（1）盛铃末期是主茎功能叶片从旺盛生理功能到衰老的转折时期;（2）从主茎上部第1叶到下部的第10叶叶绿素含量、蛋白质含量、SOD活性呈下降趋势,MDA含量、POD活性呈上升趋势;（3）盛铃期调控棉花叶片的衰老应以主茎中部轻度衰老叶片为诊断和调控主体。     

水稻早衰突变体psls1的基因定位及克隆

水稻叶片早衰直接影响作物的光合效率,减少产量,降低品质。深入研究早衰的分子机制对控制和延缓衰老具有重要意义。本文报道了一个早衰突变体psls1(premature senescence leaf with spots)的基因定位及克隆结果。突变体在发育到七叶期以后,叶片自下而上叶绿素含量下降,过氧化氢过量积累,突变体叶片逐渐黄化至枯萎;其他农艺性状如株高、分蘖数、主穗长、结实率和穗粒数也相应变差。电镜观察进一步发现,psls1衰老叶片中叶绿体降解、类囊体基粒片层模糊,嗜锇颗粒明显增多。遗传分析表明,psls1受1对隐性基因控制,利用psls1×IRAT129杂交组合F2分离群体中的1690个早衰个体,将基因PSLS1定位在第7染色体分子标记ZS-3和ZS-8之间89 kb的范围内。测序研究发现,区间内一个编码铁氧还依赖的谷氨酸合酶基因LOC_Os07g46460的第2外显子末位的G被替换为A,导致转录本的错误剪切,突变体的c DNA缺失了57 bp碱基片段。在突变体中该基因表达量下降,谷氨酸合酶活性降低,其产物谷氨酸含量显著下降、其他氨基酸代谢紊乱。水培低氮处理下可诱发突变体psls1早衰。研究结果表明,由于PSLS1突变使得谷氨酸合酶失活,氮代谢异常而导致突变体psls1早衰。     

紫叶加拿大紫荆叶色表达期相关物质的研究

以紫叶加拿大紫荆（Cercis canadensis ‘Forest pansy’）为试材,研究其在山东地区叶色表达期叶片中叶绿素、类胡萝卜素、花青苷与可溶性糖等叶色相关物质的变化。结果表明：在叶色表达期,随着天数的增加,花青苷含量总体呈下降的“M”型趋势,叶绿素和类胡萝卜素含量逐渐升高,可溶性糖含量的变化为无规律的多峰曲线;叶片中叶绿素（r=-0.681**）、类胡萝卜素（r=-0.757**）与花青苷含量均呈极显著负相关关系,可溶性糖与花青苷含量不相关;叶绿素和花青苷含量呈对数曲线关系,类胡萝卜素与花青苷含量呈指数曲线关系。     

棉花早衰、红叶茎枯病与棉花轮纹斑病间关系辨析

 系统分析了棉花早衰、红叶茎枯病与棉花轮纹斑病间的关系。棉花早衰依据其成因差异可划分为3种类型，分别为生理性早衰、病理性早衰及生理病理复合性早衰。近些年来生理病理复合性早衰在我国发生最为普遍，危害最为严重。系统分析了生理病理复合性早衰的成因及有关研究进展，在此基础上，提出适宜我国棉花生产、以“选种、壮苗、补钾、抗逆、防病”等技术为核心的棉花早衰控制技术体系。     

The Quantitative Method for Temperature Distribution of Cotton Leaves Based on Infrared Thermal Images

[Objective] The leaf temperature is an effective parameter reflecting crop physiological process and energy exchange. This study aimed to put forward a method to quantify the temperature distribution within cotton leaves. [Method] The infrared images of cotton leaves of potassium deficiency and normal cotton plants cultivated in nutrient solution were taken and studied. The irrelevant background in the infrared images was removed by using software of Stata and Surfer. The geostatistical spatial grid method was used to quantify the temperature distribution within cotton leaves. Finally, the temperature scales of different images were unified for better and faster comparison of leaf temperature differences among images. [Result] The processed image clearly reflected the differences among different treatments in leaf temperature. The mean leaf temperature, leaf vein temperature and leaf temperature outside leaf vein of potassium-deficient cotton were 0.52 ℃, 0.59 ℃ and 0.47 ℃higher than those of normal leaf, respectively. In addition, the temperature variation amplitude (0.76 ℃) and coefficient of variation (0.38%) of potassium deficient leaf were higher than those of normal leaf (0.54 ℃, 0.27%), and the temperature of the leaf vein increased the most (0.53 ℃). The analysis of the frequency distribution of temperature within the cotton leaf showed that the quartile deviation of temperature distribution of potassium-deficient leaf was 0.07 ℃ higher than that of normal leaf, indicating that the temperature distribution of normal leaf was relatively concentrated. [Conclusion] The proposed method is helpful for further monitoring of cotton growth and may guide cotton precise management.     

Effects of Verticillum Wilt on Leaf Microstructure,Photosynthesis of Cotton

[Objective] The purpose of this study was to determine the change of microstructure and photosynthetic characteristics in cotton leaves under different disease index levels (b0～b4)．[Method] Through investigation and sampling in cotton fields, five treatments were designed based on disease index levels of cotton leaf, and the leaf photosynthesis was tested by WFS-3000 and microstructure was observed with Microscope ECLIPSE Ni-E. [Result] With the increase in disease index levels, cotton leaf gradually lost green color, serious necrosis area occurred, with the leaf withered up till dried to death. The stomata of disease leaf incurred abnormally, increasing intercellular space in mesophyll tissue. Total thickness, upper and lower epidermis thickness, palisade tissue thickness and spongy tissue thickness all decreased significantly among all treatments; the trend showed as followed: total thickness was b0>b1>b2>b3>b4, upper epidermis thickness was b0>b1>b2>b3, lower epidermis thickness, palisade and spongy tissue thickness were consistent as b0>b1>b2. As a whole leaf of total thickness, upper, lower epidermis thickness, palisade and spongy tissue thickness were very significantly negatively correlated with leaf disease index levels, The correlation coefficient reached -0.6. The net photosynthesis rate, transpiration rate, stomata conductance gradually decreased and reached significant different with b0, leaf temperature increased, internal CO₂ first went up then down but no significant difference with b0. [Conclusion] The results indicated that Verticillum wilt damaged leaf microstructure and impaired photosynthesis of cotton leaf.     

超高产夏玉米花粒期不同部位叶片衰老与抗氧化酶特性

为探讨超高产玉米叶源衰老特征, 揭示其抗氧化关键酶及膜脂过氧化特性, 为玉米衰老调控和高产栽培提供依据, 本研究在大田条件下, 以我国创高产纪录的夏玉米为例, 从单株水平上对高产纪录试验(EHYR)和普通生产田(MCFF)玉米叶片衰老及抗氧化酶特性比较表明, EHYR产量达19 349 kg hm^-2, 是MCFF的2.28倍。MCFF和EHYR叶片分别在开花后30 d和50 d进入速衰期, MCFF叶片衰老比EHYR提前20 d;速衰期EHYR叶面积降幅比MCFF低5.7%。EHYR在籽粒灌浆后期, 中上部叶片净光合速率较高, 可溶性蛋白含量明显高于MCFF, 而MDA含量则维持较低水平。在叶片衰老过程中, 自开花后20 d开始, EHYR上部和中部叶片SOD活性较高, 下部叶片则以SOD、POD和CAT三者活性较高;MCFF仅中部叶片POD和CAT活性较高。EHYR叶片衰老程度与CAT活性极显著负相关, MCFF叶片衰老与SOD和POD活性显著负相关, 且二者叶片衰老进程中SOD、POD、CAT的直接作用大于间接作用。与MCFF相比, EHYR叶片除具较高SOD和POD活性外, 在籽粒灌浆后期同时保持较高CAT活性和可溶性蛋白含量是降低膜脂过氧化程度, 延缓叶片衰老的重要原因。开花后20 d是EHYR与MCFF叶片衰老出现差异的生理临界点, 因而在此时期之前调控更有利于延缓衰老。