LeCOP1LIKE基因的克隆、反义构建及微型番茄的转化

本文报告利用EST筛选结合RT-PCR的方法从番茄中克隆了LeCOP1LIKE基因的1060bpcDNA片段,并利用其非保守域构建反义RNA表达载体。利用农杆菌介导法．将LeCOP1LIKE基因的反义RNA表达载体转入微型番茄Micro-Tom．获得了10株反义LeCOP1LIKE转基因微型番茄。RT-PCR分析表明其中4个转基因株系中的LeCOP1LIKE表达被显著抑制．并发现抑制LeCOP1LIKE基因的表达导致转基因番茄的株高下降、叶片的叶绿素含量提高、果实中番茄红素含量增加,并且明显抑制转基因种子的发育。这些实验结果证明了LeCOP1LIKE基因为番茄发育过程中光形态建成的抑制因子。     

新红星苹果果实着色期几种色素含量变化及其相关性（简报）

新红星苹果果实在果皮着色前果皮内叶绿素、类胡萝卜素含量降至最低。果皮着色过程中,随果皮内花青素含量迅速增加,叶绿素及类胡萝卜素含量也出现增加趋势,果色由淡红变为深红。自然着色前在果皮伤害处理情况下可明显刺激果皮内花青素、叶绿素、类胡萝卜素含量的一致提高。     

农杆菌介导Ds转座因子转化番茄群体的构建

以番茄Micro-Tom子叶为试材,绿色荧光蛋白GFP基因为报告基因,利用农杆菌介导的遗传转化法将构建的激活表达标签pAcGFP导入番茄Micro-Tom,获得转基因番茄群体.对转化群体进行了GFP基因活性及分子检测,结果表明,GFP已经整合到番茄基因组中.以GFP基因设计引物进行PCR检测表明,阳性率为91.43%.通过Southern杂交分析,31.25%的个体为一个插入位点,56.25%为2个插入位点,T-DNA在基因组中的平均拷贝数为1.8个.     

番茄Sly-MIR167的抗冷性研究

以番茄为材料,采用Northern杂交技术,分析番茄MIR167(Sly-MIR167)在低温胁迫下的表达模式,以明确Sly-MIR167在冷胁迫下的分子调控机制,为基因工程在改良番茄品种中的实际应用提供依据。结果显示:(1)25℃下Sly-MIR167在番茄根、茎、花瓣、果实、叶片都有表达,4℃低温胁迫下的表达量均增加,表明Sly-MIR167表达受低温诱导。(2)采用农杆菌侵染构建表达载体并转化番茄获得转基因植株,冷胁迫实验结果显示:转基因植株在冷胁迫处理的生长状况明显优于对照;另外,冷胁迫下2个转基因株系(T2-5和T2-19)的最大光化学效率、叶绿素含量下降幅度明显低于野生型;脯氨酸含量高于野生型;MDA含量低于野生型,表明Sly-MIR167能够提高番茄对冷胁迫的耐受性。(3)通过miRU在线软件预测Sly-MIR167的靶基因为NF-YA1、NF-YA2,利用RT-PCR技术分析其表达下调,证明它们被MIR167负调控。     

转HAL1基因番茄的耐盐性

利用农杆菌介导的叶盘法,把HAL1 基因转入番茄,Southern杂交检测得到转基因植株.耐盐实验表明, T1代转基因番茄在150 mmol/L的NaCl胁迫下仍有43%的发芽率,200 mmol/L的NaCl胁迫下发芽率为6%,而对照种子在100和150 mmol/L的NaCl胁迫下发芽率分别为11.0%和0.转基因番茄的电解质相对外渗率小于对照,而根冠比和叶绿素含量大于对照,转HAL1基因显著提高了番茄的耐盐性.盐胁迫下Na 、K 的累积状况表明,转基因番茄根、茎、叶的K /Na 均有所提高,根系的SK/Na增大,茎、叶的RSK/Na和RLK/Na减小,说明根系对K /Na 离子的选择吸收和运输能力加强.不但选择吸收K /Na ,而且表现出整株水平上的有利于耐盐的K /Na 区域化分配.     

UGPase和反义4CL基因对转基因烟草纤维素和木质素合成的调控

用根癌农杆菌介导法将源于紫穗槐的尿苷二磷酸葡萄糖焦磷酸化酶（UGPase）基因、反义4-香豆酸辅酶A连接酶（4CL）基因以及两者的双价基因分别转移至烟草中。PCR和Southern杂交检测证实外源基因已整合到转基因烟草基因组中。测定全纤维素和Klason木质素含量的结果显示,增强UGPase基因的表达可提高转基因植株的纤维素含量,但对木质素含量没有影响;抑制4CL基因的表达可显著降低转基因植株的木质素含量,但对纤维素含量没有影响;转移双价基因的转基因植株中纤维素含量增加而木质素含量降低。     

OsPT6基因过表达对低磷条件下菜用大豆结瘤及固氮的影响

采用砂培方法,以转OsPT6基因的菜用大豆(T3株系)与其非转基因(NT)受体品种为实验材料,研究了两者在低磷条件下的生长发育指标,植株有效磷、全磷、全氮、豆血红蛋白和籽粒蛋白质含量以及谷氨酰胺合成酶活性的差异,并对植株结瘤及固氮相关基因表达进行检测,为阐明转OsPT6基因菜用大豆在低磷条件下结瘤及固氮相关机理提供理论依据。结果显示:(1)转基因植株的株高、茎粗、花数和荚数、根瘤数均显著高于NT植株。(2)转基因植株根、茎、叶及根瘤中有效磷,全株总磷、总氮含量,根瘤中的豆血红蛋白含量、功能叶片中谷氨酰胺合成酶的活性和籽粒蛋白质含量均显著高于NT植株。(3)相关性分析表明,豆血红蛋白含量、谷氨酰胺合成酶活性、总磷、总氮含量4个指标间均呈显著正相关关系。(4)GmENOD40a、GmENOD40b、GmGS1β1、GmGS1β2基因在转基因植株中的表达量显著高于NT植株。研究表明,OsPT6基因过表达增强了菜用大豆在低磷条件下的结瘤及固氮能力,该研究结果为进一步研究其调控机理奠定了基础。     

转星星草金属硫蛋白基因烟草的获得及其抗重金属镉能力分析

用农杆菌介导法将星星草金属硫蛋白基因（MD转化烟草,PCR及PCR—Southern检测的结果表明,该基因已导入烟草中。Real-TimePCR检测显示,该基因在转基因后代中的转录水平高于非转基因植株。Cd2＋胁迫下,转基因植株能够正常生长,鲜重、株高、叶绿素含量、Cd2＋含量和SOD活性均高于非转基因植株,表明MT基因的过量表达可提高转基因烟草的抗Cd2＋能力。     

紫色马铃薯花青素StAN1基因的克隆及功能分析

MYB是植物花青素合成代谢途径中最主要的转录因子。该研究以紫色马铃薯B-5为试验材料,通过同源克隆技术,克隆了马铃薯花青素StAN1基因,并对StAN1基因的表达、遗传转化及其转基因烟草的花青素含量进行分析。结果表明:(1)StAN1基因全长774bp,编码了258个氨基酸;系统进化树分析发现,StAN1与辣椒、茄子、芦竹的亲缘关系最近,与矮牵牛、苹果等的亲缘关系最远。(2)荧光定量PCR分析显示,StAN1基因在马铃薯不同组织均有表达,其表达量从小到大依次为:根叶茎匍匐茎薯皮薯肉。(3)成功构建了表达载体pJAM1502-AN1,并经农杆菌(Gv3101)转化获得根、茎、叶片及叶脉均为紫红色的转StAN1基因烟草,PCR鉴定表明目的基因StAN1已成功转入烟草中。(4)花青素含量分析表明,野生型烟草叶片中花青素含量为2mg/g,叶片颜色为绿色,而转StAN1基因烟草叶片花青素含量达20mg/g,叶片颜色变成了紫红色。     

Changes in growth and photosynthetic characteristics of Ocimum sanctum under growth regulator treatments

A field experiment was conducted to investigate the effect of growth regulators on growth characteristics such as root length, shoot length, total leaf area, number of inflorescence per plant, number of flower per inflorescence, whole plant fresh weight and whole plant dry weight. Photosynthetic characteristics were also analyzed based on the same experiment. For this, various photosynthetic pigment contents such as chlorophyll, carotenoid, anthocyanin and xanthophyll content were calculated. The conventional growth regulator abscisic acid (ABA) and non-conventional growth regulator triazole compound paclobutrazol (PBZ) were used. Root length increased due to growth regulator treatment, but shoot length decreased. Leaf area was decreased due to growth regulator treatment. The number of inflorescence increased in ABA treated plants, but it was decreased in PBZ treated plants. In ABA treated plants, the number of flowers per inflorescence was increased. In PBZ treated plants the number of inflorescence was reduced. The whole plant fresh weight (FW) and dry weight (DW) were increased in ABA and PBZ treated plants. There was an increase in chlorophyll content in growth regulator treated plants compared to control, and it was more in PBZ treated plants. The carotenoid content was also increased in ABA and PBZ treated plants.     

Changes in growth and photosynthetic characteristics of Ocimum sanctum under growth regulator treatments

A field experiment was conducted to investigate the effect of growth regulators on growth characteristics such as root length,shoot length,total leaf area,number of inflorescence per plant,number of flower per inflorescence,whole plant fresh weight and whole plant dry weight.Photosynthetic characteristics were also analyzed based on the same experiment.For this,various photosynthetic pigment contents such as chlorophyll,carotenoid,anthocyanin and xanthophyll content were calculated.The conventional growth regulator abscisic acid (ABA) and non-conventional growth regulator triazole compound paclobutrazol (PBZ) were used.Root length increased due to growth regulator treatment,but shoot length decreased.Leaf area was decreased due to growth regulator treatment.The number of inflorescence increased in ABA treated plants,but it was decreased in PBZ treated plants.In ABA treated plants,the number of flowers per inflorescence was increased.In PBZ treated plants the number of inflorescence was reduced.The whole plant fresh weight (FW) and dry weight (DW) were increased in ABA and PBZ treated plants.There was an increase in chlorophyll content in growth regulator treated plants compared to control,and it was more in PBZ treated plants.The carotenoid content was also increased in ABA and PBZ treated plants.     

植物叶片花青素的光破坏防御机制研究进展

植物花青素广泛分布在植物的根、茎、叶、花和果实等器官中,是植物形态建成过程中或响应逆境而产生的一种次生代谢物质.植物叶片中的花青素具有特殊的化学结构和光谱特性,在光破坏防御机制方面发挥了重要的作用,已经成为植物光合生理生态的研究热点.本文综述了近年来植物叶片花青素与光合作用的研究进展,从叶片花青素的分布、光谱特性及其与光合色素的关系等方面说明花青素对植物光合作用的影响,重点介绍了叶片花青素通过光吸收、抗氧化剂和渗透调节等在植物光破坏防御机制方面的作用,展望了今后的主要研究方向     

Expression of isopentenyl transferase gene (<Emphasis Type="Italic">ipt</Emphasis>) in leaf and stem delayed leaf senescence without affecting root growth

A cytokinin biosynthetic gene encoding isopentenyl transferase (ipt) was cloned with its native promoter from Agrobacterium tumefaciens and introduced into tobacco plants. Indolebutyric acid was applied in rooting medium and morphologically normal transgenic tobacco plants were regenerated. Genetic analysis of self-fertilized progeny showed that a single copy of intact ipt gene had been integrated, and T₂ progeny had become homozygous for the transgene. Stable inheritance of the intact ipt gene in T₂ progeny was verified by Southern hybridization. Northern blot hybridization revealed that the expression of this ipt gene was confined in leaves and stems but undetectable in roots of the transgenic plants. Endogenous cytokinin levels in the leaves and stems of the transgenic tobaccos were two to threefold higher than that of control, but in roots, both the transgenic and control tobaccos had similar cytokinin levels. The elevated cytokinin levels in the transgenic tobacco leaves resulted in delayed leaf senescence in terms of chlorophyll content without affecting the net photosynthetic rate. The root growth and morphology of the plant were not affected in the transgenic tobacco.     

Maternal flower color,ultraviolet protection,and germination in Ipomopsis aggregata (Polemoniaceae)

Transgenerational interactions between flower color, seed quality, and seedling performance have rarely been investigated. The ecological model, Ipomopsis aggregata, is a great candidate for examining the maternal effects of flower color because it is a mostly scarlet-flowering plant which shows color polymorphism within natural populations. Anthocyanin, the red flavonoid pigment which gives these flowers color, has been shown to act as an ultraviolet (UV) protectant by shielding chloroplasts and acting as an antioxidant. This study was conducted on scarlet- and fuchsia-flowering maternal plants and their seeds from natural populations in Colorado. Dark-flowering (scarlet) maternal plants from these populations had consistently higher foliar anthocyanin content, photosystem efficiency, and chlorophyll content than light-flowering (fuchsia) plants over a 3-year period in the field. Seeds from a subset of these maternal plants were counted, weighed, and germinated in a growth chamber. Photosystem efficiency, vegetative anthocyanin content, chlorophyll content, and biomass were measured on germinated seedlings after the germination census was completed. Dark-flowering maternal plants yielded seeds and seedlings with higher biomass than light-flowering ones. Seeds from dark-flowering maternal plants also germinated faster than those from light-flowering maternal plants and seedlings had higher vegetative anthocyanin content. The hereditary nature of anthocyanin content thus suggests that higher anthocyanin levels (both floral and vegetative) are potentially linked to measures of fitness such as increased seed weight, germination rate, and seedling biomass. These data suggest that UV protection provided by anthocyanins potentially increases the realized fitness of maternal plants, thereby influencing life history.     

苗期遮光光质对生姜光合及生长的影响

以不同颜色塑料薄膜为遮光材料,研究了苗期遮光光质对生姜生长及光合作用的影响.结果表明:幼苗覆膜期,生姜叶片叶绿素含量以蓝膜及绿膜处理较高,白膜次之,红膜较低;叶片P_n则以绿膜处理较高,为14.9 μmol·m^-2·s^-1（第4叶）,分别较白膜、红膜及蓝膜提高5.7%、10.4%和18.3%.旺盛生长期撤膜后,P_n较幼苗期升高,但处理间的变化趋势与幼苗期相似;新生叶片叶绿素含量除红膜处理较低外,其它处理无显著差异,但下位叶片叶绿素含量则以蓝膜和红膜处理显著低于绿膜和白膜处理.蓝膜处理生姜植株茎秆增高、变细,分枝数较少;绿膜处理植株根、茎、叶及根茎鲜质量较高,白膜、红膜及蓝膜处理依次降低,收获时,其产量分别达57 000、53 709、51 487和48 712 kg·hm^-2.说明生姜苗期采用绿膜遮光,可增强叶片光合作用,促进植株生长,提高生姜产量.     

叶面喷施茉莉酸甲酯对马铃薯微型薯生长及产量的影响

以‘云薯505’马铃薯(Solanum tuberosum ‘Yunshu 505’)为材料,测定马铃薯块茎发育初期四个阶段茉莉酸含量,并以叶面喷施方式,研究茉莉酸甲酯对马铃薯生长和块茎产量的影响。结果表明,马铃薯块茎膨大过程中,茉莉酸的积累水平不断升高。在微型薯生产中,使用100 μmol·L^-1茉莉酸甲酯在结薯期以不同频率喷施叶面,测量并统计植株、块茎性状及产量变化。结果表明,与对照(CK)相比,1次·d^-1处理茎粗增加36.1%,2次·d^-1处理的叶绿素含量降低20.1%。此外,植株的叶色、茎色、花色、株型等生长性状及块茎大小整齐度、薯形、皮色、肉色、薯皮类型、芽眼深浅、裂薯率、大薯空心率等块茎性状在各组间没有显著差异。2次·d^-1、1次·d^-1、1次·2d^-1、CK四种处理的植株存活率分别为45.57%、100.00%、100.00%、87.29%;前三种喷施频率处理折合产量较CK分别增加-15.61%、8.77%、12.11%。综合分析,马铃薯在块茎形成初期茉莉酸积累水平不断升高,以1次·2d^-1频率叶面喷施100 μmol·L^-1茉莉酸甲酯,马铃薯微型薯的产量增加最大且不影响生长。     

Genetic engineering of the anthocyanin biosynthetic pathway with flavonoid-3′,5′-hydroxylase: specific switching of the pathway in petunia

Flavonoid-3',5'-hydroxylase (F3'5'H) is the key enzyme in the synthesis of 3',5'-hydroxylated anthocyanins, which are generally required for the expression of blue or purple flower color. It has been predicted that the introduction of this enzyme into a plant species that lacks it would enable the production of blue or purple flowers by altering the anthocyanin composition. We present here the results of the genetic engineering of petunia flower color, pigmentation patterns and anthocyanin composition with sense or antisense constructs of the F3'5'H gene under the control of the CaMV 35S promoter. When sense constructs were introduced into pink flower varieties that are deficient in the enzyme, transgenic plants showed flower color changes from pink to magenta along with changes in anthocyanin composition. Some transgenic plants showed novel pigmentation patterns, e.g. a star-shaped pattern. When sense constructs were introduced into blue flower petunia varieties, the flower color of the transgenic plants changed from deep blue to pale blue or even pale pink. Pigment composition analysis of the transgenic plants suggested that the F3'5'H transgene not only created or inhibited the biosynthetic pathway to 3',5'-hydroxylated anthocyanins but switched the pathway to 3',5'-hydroxylated or 3'-hydroxylated anthocyanins.