GA2ox8基因过量表达诱导蓝光下拟南芥光形态建成

检测不同光照强度蓝光下,过量表达GA2ox8基因的转基因植株光形态建成表型的结果表明,突变体比各自的母本的下胚轴短,茎尖角度和子叶张开度较大,花青素和叶绿素的含量较高,并且其差异与GA2ox8基因的表达量呈正相关.RT-PCR检测光调节基因表达水平的结果显示,暗培养条件下其突变体幼苗中的水平比各自的母本高,蓝光下35S::GFP-GA2ox8-1.35S::GFP-GA2ox8-8与母本co1-4之间差异不明显,但scc7-D中的水平比母本crylcry2高.这似乎说明,GA2ox8基因过量表达可诱导蓝光下拟南芥幼苗光形态建成.     

GA20x8基因过量表达诱导蓝光下拟南芥光形态建成

检测不同光照强度蓝光下,过量表达GA20x8基因的转基因植株光形态建成表型的结果表明,突变体比各自的母本的下胚轴短,茎尖角度和子叶张开度较大,花青素和叶绿素的含量较高,并且其差异与GA20x8基因的表达量呈正相关。RT—PCR检测光调节基因表达水平的结果显示,暗培养条件下其突变体幼苗中的水平比各自的母本高,蓝光下35S：：GFP—GA20x8-1,35S：：GFP—GA20x8—8与母本col-4之间差异不明显,但scc7-D中的水平比母本crylcry2高。这似乎说明,GA20x8基因过量表达可诱导蓝光下拟南芥幼苗光形态建成。     

CONSTANS LIKE 7参与调控拟南芥生物钟

植物的生物钟节律可以被环境中的光/黑暗以及冷/热循环所诱导,并使其与环境同步。植物生物钟由输入途径、中央振荡器、输出途径组成。目前对植物生物钟的研究已经揭示生物钟最基本的组成,但是关于生物钟的运作机理及网络还需要进一步研究。CONSTANS LIKE 7(COL7)是CONSTANS(CO)的家族基因。以拟南芥野生型(wild type,WT)、突变体col7以及COL7过量表达转基因株系COL7-OX-10和COL7-OX-11为材料,利用定量PCR、叶片运动等方法,分析COL7是否受生物钟调控以及COL7是否参与调控生物。实验结果显示:COL7不仅受生物钟调控,同时也参与调控生物钟。     

光质对水稻幼苗初级氮同化的影响

用滤光膜过滤蓝色或红色荧光灯,得到纯的蓝光和红光,以白光为对照,研究不同光质对水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）幼苗初级氮同化的影响。结果表明：蓝光促进水稻黄化幼苗吸收ＮＯ＾－３含量,并促进ＮＲ（硝酸还原酶）的诱导。在蓝光下生长５￣７ｄ的幼苗的ＮＲ、ＮＩＲ（亚硝酸还原酶）、ＧＳ（谷氨酰胺合成酶）和ＧＯＧＡＴ（谷氨酸合酶）活性均高于白光下生长的,但第１０天以后,白光下生长的幼苗酶活性最高。与白光     

NaCl胁迫对姜黄组培苗生理特性的影响

以姜黄组培苗为试验材料,在继代培养基中分别添加0、25、50、75、100、125、150mmol·L-1NaCl进行盐胁迫处理,研究NaCl胁迫下姜黄幼苗生长和生理特性的变化。结果表明:NaCl胁迫下,姜黄组培苗的脯氨酸和丙二醛含量逐渐上升,而叶绿素和类胡萝卜素含量、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性逐渐下降。25 mmol·L-1NaCl时姜黄幼苗的基径、株高和鲜重与对照差异不显著,50 mmol·L-1NaCl时则显著下降;表明姜黄组培苗的生长可以耐受25~50 mmol·L-1NaCl胁迫。随着NaCl浓度增加,姜黄幼苗的可溶性糖和蛋白质含量、过氧化物酶(POD)活性先升高后降低,75 mmol·L-1NaCl时均达到最大值。50~100 mmol·L-1NaCl胁迫下姜黄叶片的苯丙氨酸解氨酶(PAL)活性和姜黄素含量得到显著提高,表明50~100 mmol·L-1NaCl胁迫对姜黄素的合成有促进作用。可见,25~50 mmol·L-1NaCl能够促进姜黄植株的生长,而75 mmol·L-1NaCl显著提高次生代谢产物姜黄素的含量。     

钙与植物乙烯反应的关系研究

研究了Ca2 对番茄 (LycopersiconesculentumMillcv.Lichun)黄化幼苗乙烯反应的影响。通过测定不同Ca2 浓度条件下番茄黄化幼苗的“三重反应”、内源乙烯释放量、乙烯受体基因NEVER_RIPE(NR)表达量及胞内CaM含量的变化 ,结果发现 ,随着培养基中Ca2 浓度从 0mmol/L增加到 3.8mmol/L ,番茄黄化幼苗的“三重反应”表型明显增强 ,内源乙烯释放量、NR基因的表达量及胞内CaM的含量都有不同程度的增加 ;当Ca2 浓度由 3.8mmol/L进一步增加到 10mmol/L时 ,番茄黄化幼苗“三重反应”表型受到抑制 ,内源乙烯释放量、NR基因的表达量及胞内CaM的含量都有所下降。因此 ,Ca2 对番茄黄化幼苗“三重反应”的影响与Ca2 调节内源乙烯合成和乙烯受体基因的表达有关 ,而且Ca2 可能是通过CaM含量的变化来调节乙烯作用的     

转金属硫蛋白基因(MT_1)烟草耐NaCl胁迫能力

为明确柽柳(Tamarix sp.)金属硫蛋白(MT1)基因过量表达对提高植物耐NaCl能力的作用,对转MT1因烟草进行分子检测和生理特性分析,结果表明具有卡那霉素抗性的转基因植株经RT-PCR Southern杂交均表现为阳性,说明外源MT1基因已整合到烟草基因组,并且得到了表达。金属硫蛋白基因的过量表达提高了转基因烟草植株的耐NaCl能力,表现为在含有150mmol/L和300mmol/L NaCl的MS培养基上,转基因植株的株高和鲜重均明显优于非转基因株系;在生理性状上表现为转基因植株丙二醛(MDA)含量明显低于非转基因株系,而超氧化物歧化酶(SOD)、过氧化物酶(POD)活性比非转基因株系明显增加。     

钙与植物乙烯反应的关系研究

研究了Ca2+ 对番茄(Lycopersicon esculentum Mill cv. Lichun)黄化幼苗乙烯反应的影响.通过测定不同Ca2+ 浓度条件下番茄黄化幼苗的"三重反应"、内源乙烯释放量、乙烯受体基因NEVER-RIPE(NR)表达量及胞内CaM含量的变化,结果发现,随着培养基中Ca2+ 浓度从0 mmol/L增加到3.8 mmol/L,番茄黄化幼苗的"三重反应"表型明显增强,内源乙烯释放量、NR基因的表达量及胞内CaM的含量都有不同程度的增加;当Ca2+ 浓度由3.8 mmol/L进一步增加到10 mmol/L时,番茄黄化幼苗"三重反应"表型受到抑制,内源乙烯释放量、 NR基因的表达量及胞内CaM的含量都有所下降.因此,Ca2+ 对番茄黄化幼苗"三重反应"的影响与Ca2+ 调节内源乙烯合成和乙烯受体基因的表达有关,而且Ca2+ 可能是通过CaM含量的变化来调节乙烯作用的.     

蓝光调节高粱突变体har1 幼苗的去黄化反应

以航空诱变高粱突变体har1为材料, 对其幼苗去黄化过程进行研究。萌发的种子在远红光下预培养6小时后, 置于12小时蓝光/12小时黑暗条件下培养。测量幼苗的各器官伸长, 结果表明, 与野生型R111相比, har1的胚芽鞘、中胚轴、第一叶鞘以及第二叶鞘的伸长均受到蓝光的明显抑制, 而蓝光对叶片生长影响不明显。3天龄har1黄化苗在连续蓝光下中胚轴花色素苷的积累明显增高, 红光和远红光无此效应。此外, 蓝光促进har1叶片叶绿体发育, 且在蓝光照射24小时后叶片中叶绿素含量升高。Western blot检测结果显示, 7天龄R111和har1幼苗隐花色素SbCRY1b蛋白水平呈现蓝光下低、黑暗中高的变化趋势, har1的SbCRY1b蛋白水平在黑暗中高于R111。研究结果表明, 高粱har1在去黄化过程中具有蓝光超敏感表型,SbCRY1b的作用值得进一步深入研究。     

蓝光调节高粱突变体har1幼苗的去黄化反应

以航空诱变高粱突变体har1为材料,对其幼苗去黄化过程进行研究。萌发的种子在远红光下预培养6小时后,置于12小时蓝光/2小时黑暗条件下培养。测量幼苗的各器官伸长,结果表明,与野生型R111相比,harl的胚芽鞘、中胚轴、第一叶鞘以及第二叶鞘的伸长均受到蓝光的明显抑制,而蓝光对叶片生长影响不明显。3天龄har1黄化苗在连续蓝光下中胚轴花色素苷的积累明显增高,红光和远红光无此效应。此外,蓝光促进har1叶片叶绿体发育,且在蓝光照射24小时后叶片中叶绿素含量升高。Westernblot检测结果显示,7天龄R111和har1幼苗隐花色素SbCRY1b蛋白水平呈现蓝光下低、黑暗中高的变化趋势,har1的SbCRY1b蛋白水平在黑暗中高于R111。研究结果表明,高粱har1在去黄化过程中具有蓝光超敏感表型,SbCRY1b的作用值得进一步深入研究。     

转GFP::cDNA基因拟南芥筛选及相关基因的功能分析

本研究以随机GFP::cDNA融合基因转基因拟南芥为材料,筛选到在细胞核或在细胞核和细胞质中均表达GFP信号的转基因株系58个。对这些转基因株系中的cDNA插入片段进行克隆,获得4株插入片段能按原初编码框进行编码的转基因株系。对插入片段为编码富含甘氨酸蛋白AtGRP8 C-末端(富含甘氨酸的结构域)的转基因株系R2的表型分析发现,连续白光、红光或蓝光下其幼苗的下胚轴比野生型的要短,且较低光照强度白光(低于100μmol m-2s-1)、蓝光(低于75μmol m-2s-1)下的差异更加明显,但是黑暗中其幼苗的下胚轴与野生型相比无明显差异,表明AtGRP8蛋白可能通过其C-末端功能域参与调控拟南芥的光形态建成反应。     

蔗糖调节拟南芥花青素的生物合成

为了探讨糖在花青素合成过程中的调节作用,采用蔗糖和其代谢糖（葡萄糖 和果糖）组合处理拟南芥幼苗.实验结果表明,60 mmol/L蔗糖处理显著提高拟南芥 幼苗的花青素、还原糖含量,并上调花青素合成相关基因（CHS, FLS-1, DFR, LDOX, BANYULS）的转录,对叶绿素含量和UGT78D2基因的转录无影响;20 mmol/L 葡萄糖+20 mmol/L果糖处理,对花青素、叶绿素和还原糖的含量无影响,对花青素 合成相关基因转录影响不一;20 mmol/L蔗糖+20 mmol/L葡萄糖+20 mmol/L果糖处 理后,花青素和还原糖含量介于前两个处理之间,也上调花青素合成相关基因的转 录;但和蔗糖处理组相比,上调UGT78D2基因转录,下调FLS-1基因转录.在不同处 理组之间,花青素含量变化和还原糖含量变化趋势相同,有可能糖在调节花青素 合成的同时也调节还原糖含量.因此,蔗糖既可以通过蔗糖特异信号途径,也可以 和其代谢糖通过其他途径共同调节拟南芥花青素的生物合成.     

6-BA预处理对低温弱光胁迫下辣椒幼苗叶绿素a荧光参数和膜脂过氧化的

研究6-BA预处理对低温（15℃/5℃）弱光（100μmol·m^-2·s^-1）下辣椒品种‘湘研16号’幼苗叶片中叶绿素a荧光参数和膜脂过氧化影响。结果表明,0．08mmol·L^-16-BA减缓低温弱光下辣椒幼苗光系统II（PSII）最大光化学效率（Fv/Fm）、PSH实际光化学效率（ΦPSII）和PSII反应中心光能捕获效率（Fv’/Fm’）的下降以及叶片膜透性和丙二醛（malondialdehyde,MDA）含量的增加,但是提高了超氧化物歧化酶（superoxidedismutase,SCD）、过氧化物酶（peroxidase,POD）、过氧化氢酶（catalase,CAT）以及抗坏血酸过氧化物酶（ascorbate peroxidase,APX）的活性。     

Manipulation of the blue light photoreceptor cryptochrome 2 in tomato affects vegetative development, flowering time, and fruit antioxidant content

Cryptochromes are blue light photoreceptors found in plants, bacteria, and animals. In Arabidopsis, cryptochrome 2 (cry2) is involved primarily in the control of flowering time and in photomorphogenesis under low-fluence light. No data on the function of cry2 are available in plants, apart from Arabidopsis (Arabidopsis thaliana). Expression of the tomato (Solanum lycopersicum) CRY2 gene was altered through a combination of transgenic overexpression and virus-induced gene silencing. Tomato CRY2 overexpressors show phenotypes similar to but distinct from their Arabidopsis counterparts (hypocotyl and internode shortening under both low- and high-fluence blue light), but also several novel ones, including a high-pigment phenotype, resulting in overproduction of anthocyanins and chlorophyll in leaves and of flavonoids and lycopene in fruits. The accumulation of lycopene in fruits is accompanied by the decreased expression of lycopene beta-cyclase genes. CRY2 overexpression causes an unexpected delay in flowering, observed under both short- and long-day conditions, and an increased outgrowth of axillary branches. Virus-induced gene silencing of CRY2 results in a reversion of leaf anthocyanin accumulation, of internode shortening, and of late flowering in CRY2-overexpressing plants, whereas in wild-type plants it causes a minor internode elongation.     

Arabidopsis cryptochrome 1 is a soluble protein mediating blue light-dependent regulation of plant growth and development

Cryptochrome 1 (CRY1) is a flavin-type blue light receptor of Arabidopsis thaliana which mediates inhibition of hypocotyl elongation. In the work described in this report it is demonstrated that CRY1 is a soluble protein expressed in both young seedlings grown either in the dark or under light, and in different organs of adult plants. The functional role of CRY1 was further investigated using transgenic Arabidopsis plants overexpressing CRY1. It is demonstrated that overexpression of CRY1 resulted in hypersensitivity to blue, UV-A, and green light for the inhibition of hypocotyl elongation response. Transgenic plants overexpressing CRY1 also exhibited a dwarf phenotype with reduced size in almost every organ. This was in keeping with the previous observation of reciprocal alterations found in hy4 mutant plants and is consistent with a hypothesis that CRY1 mediates a light-dependent process resulting in a general inhibitory effect on plant growth. In addition, transgenic plants overexpressing CRY1 showed increased anthocyanin accumulation in response to blue, UV-A, and green light in a fluence rate-dependent manner. This increase in anthocyanin accumulation in transgenic plants was shown to be concomitant with increased blue light-induction of CHS gene expression. It is concluded that CRY1 is a photoreceptor mediating blue light-dependent regulation of gene expression in addition to its affect on plant growth.     

Influence of Light of Different Spectral Compositions on Growth and Metabolism of Citrus Seedlings

Sour orange (Citrus aurantium L.) seedlings grown for six months under covers transmitting light of different spectral composition, were compared with others grown under a white cover (control) and outside in full daylight. The intensity of transmitted light was equalized under all covers and attained only 20% of full daylight. Seedlings grown in daylight were shorter, had more internodes, smaller leaves, less chlorophyll and more ascorbic acid than the others. Blue + far-red covers (no transmission between 560–700 nm) enhanced seedling length, the protein and chlorophyll content and peroxidase activity of leaves. When also the wave-range above 700 nm was cut out (blue) seedlings were the shortest, and leaves had very high protein and chlorophyll content, but much less ascorbic acid and lower peroxidase activity. Red + far-red covers (no transmission below 500 nm) enhanced seedling length more than blue + far-red; leaves contained as much protein as control, but had relatively high chlorophyll and peroxidase activity. Ascorbic acid was as low as in blue light.     

外源钙降低拟南芥幼苗盐害效应

用5mmol／L CaCl2喷叶后的拟南芥幼苗,在100mmol／L NaCl胁迫下,SOD、CAT和POD的活性以及叶绿素含量降低幅度明显低于对照,MDA含量增加幅度显著低于对照,由此表明钙与拟南芥幼苗盐害效应的降低有关。     

Molecular cloning and expression analysis of a CONSTANS homologue,PnCOL1, from Pharbitis nil

The Arabidopsis CONSTANS (CO) gene is a key regulator of the long day (LD)-dependent flowering pathway and two CO homologous genes COL1 and COL2 are involved in the regulation of the circadian rhythm. In order to understand the role of CO and COL in short-day plants, a CO homologue, PnCOL1, was isolated and characterized from Japanese morning glory (Pharbitis nil). The deduced PnCOL1 protein of 386 amino acid residues contained two putative zinc finger motifs at the N-terminal region and a conserved CCT domain at the C-terminal region. The deduced amino acid sequence of PnCOL1 was 34% identical to that of PnCO, but 32%, 34%, and 34% identical to those of CO, COL1, and COL2, respectively. Expression of PnCOL1 was barely detected in the cotyledons of plants grown under continuous light (CL), but highly expressed in the cotyledons of plants grown under SD. Expression of PnCOL1 showed a pattern of circadian rhythm as well as daily oscillation. The overexpression of PnCOL1 by a 35S promoter did not overcome the late-flowering phenotype of Arabidopsis co mutants. The results provided in this study suggest that PnCOL1 may have a role in the circadian rhythm in Pharbitis nil.     

光处理对白头翁愈伤组织生长及代谢产物合成的影响

以白头翁试管苗叶片为外植体,于MS＋TDZ0.2 mg/L＋2,4-D 0.2 mg/L上进行愈伤组织诱导和增殖,进行不同光质照射,观察光处理下白头翁愈伤组织的生长和代谢产物合成情况.结果表明：黄光能提高愈伤组织的诱导率,产生的愈伤组织质地最好;光质对愈伤组织的鲜重增殖倍数依次为：黄（16.69）〉 红（11.41）〉 绿（8.97）〉 白（6.74）〉蓝（5.97）,延长光照时间会促进细胞生长;与白光相比,黄光和红光能显著促进干物质的积累,而蓝光明显抑制,绿光与白光两处理之间差异不明显.自然光照下,白头翁叶片中代谢产物极少,而黄、蓝和绿光均诱导合成了包括皂苷在内的数种代谢产物,而红光处理同白光一样只诱导出了一种代谢产物.由此可见,光处理可以调控白头翁愈伤组织的生长状态及皂苷等次生代谢物的积累.     

超表达AVP1基因提高转基因百脉根的耐盐性和抗旱性

本研究以超表达拟南芥液泡膜H＋-焦磷酸酶编码基因AVPI的转基因百脉根为材料,对其耐盐性和抗旱性进行了检测。结果显示：在200mmol·L^-1 NaCl下处理或自然干旱7d后,转基因植株的生长虽然受到抑制,但受抑程度明显低于野生型植株,前者叶片相对含水量比后者分别高18％和14％,净光合速率分别高20％和21％,而MDA含量则分别低35％和27％,相对质膜透性分别低28％和27％。此外,随着盐和干旱胁迫的加剧,与野生型植株相比,转基因植株体内积累了更多Na＋、K＋和Ca2＋。以上结果表明,AVPI基因的超表达可能提高了百脉根细胞Na＋区域化能力,既减轻了过量Na＋对细胞质的毒害作用,也提高了植株的渗透调节能力,从而增强了百脉根的耐盐性和抗旱性。