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1.
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2.
Major progress has been made over the last few years in the identification and regulation of tomato ripening genes. At least 25 genes showing elevated expression during ripening have been cloned and several, including polygalacturonase, which modifies fruit textures, have been shown to be ripening-specific. In addition, genes have been cloned for ACC synthase and ACC oxidase, which control the synthesis of ethylene, which plays a critical role in ripening. Inhibition of expression of polygalacturonase, pectinesterase, ACC synthase, ACC oxidase and phytoene synthase has been achieved in transgenic plants, using antisense technology. The expression of several genes has also been inhibited by sense gene suppression. New traits caused by these transgenes are stably inherited. Antisense tomatoes with reduced polygalacturonase have improved textural qualities which are being exploited commercially for the fresh and processed markets. Overexpression of phytoene synthase has been shown to restore carotenoid production in the yellow flesh mutant and can be used to enhance colour in other cultivars. Antisense fruit in which ACC synthase or ACC oxidase are inhibited show slower ripening and reduced over-ripening. ACC oxidase antisense genes have also been shown to delay leaf senescence. It is to be expected that further genes determining other quality traits will be identified and manipulated soon.  相似文献   

3.
Summary The common cultivated tomato (Lycopersicon esculentum Mill.) provides a major focus for improvement of crop quality through genetic engineering. Identification of ripening-related cDNAs has enabled the modification of specific aspects of ripening by manipulating gene expression in transgenic plants. By utilizing antisense RNA to modify expression of ripening genes, we have inhibited the production of the cell wall-metabolising enzymes polygalacturonase and pectinesterase and created transgenic plants that contain, effectively, single, targeted mutations affecting these genes. Furthermore, this approach has been used with previously unidentified cDNA clones to enable both functional identification and manipulation of genes involved in ethylene production (ACC oxidase) and carotenoid biosynthesis (phytoene synthase). The use of antisense RNA targeted to specific genes to alter ripening phenotypes and improve commercial utility of fruit by affecting shelf-life, processing characteristics and nutritional content is discussed.We have used the extreme ripening-impaired mutant, ripening inhibitor (rin) to identify additional genes implicated in the ripening process. This approach has resulted in the cloning of several novel ripening-related mRNAs which are now being studied by antisense experiments. This may enable identification and manipulation of additional genes involved in processes such as softening, flavour and aroma generation and susceptibility to pathogens.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - PE pectinesterase - PG polygalacturonase - SAM S-adenosyl methionine - SARs scaffold attachment regions  相似文献   

4.
The specific ACC oxidase 1 gene fragment of tomato was cloned and prepared for a probe which was used to hybridize with total RNAs that were extracted from the materials treated with ethylene and different environmental stresses, such as drought, water recovery, flood, wound and temperature, respectively. Northern blot analysis indicated that the expression of LeACO1 was restrained by drought, while strongly induced by water recovery and exogenous ethylene. After treated by flood stress, the expression level of I_eACO1 in leaves had no distinctively changes, while its expression level in stems was rapidly increased. Moreover, the expression of LeACO1 in tomato stems could be induced at the temperature of 37 degree. In addition, wounding had no influence on the expression of LeACO1.  相似文献   

5.
Flowers of Dendrobium cv. Kenny were hand-pollinated using pollinia from cv. Sakura. This resulted in a large increase in flower ethylene emission and rapid perianth (tepal) senescence. The increase in flower ethylene emission was correlated in time with an increase in ethylene emitted by the column (the fused stigma, style and stamens) plus the ovary. No ethylene emission was observed from perianth parts that were isolated at various periods after pollination. The increased ethylene emission by the column plus ovary was correlated with an increase in ACC synthase and ACC oxidase activity in these flower parts. The perianth parts, in contrast, only showed an increase in ACC oxidase activity, following pollination. The data show that pollination-induced early perianth senescence in Dendrobium is mediated by increased ethylene biosynthesis by the column + ovary, and not due to increased ethylene biosynthesis in the perianth parts. Apparently, ethylene synthesized in the gynoecium diffuses to the perianth parts where it induces senescence. The data are very similar to those found previously in pollinated Phaleanopsis orchids and in emasculated Cymbidium orchids, with the exception that ethylene was emitted from the tepals of these two orchids and not from Dendrobium.  相似文献   

6.
Fruit of cv. Gros Michel banana were treated with 1-MCP (1000 nL L−1 for 4 h at 25 °C) and then packed in non-perforated polyethylene (PE) bags for modified atmosphere storage (MAP). The bags were placed in corrugated cardboard boxes and stored at 14 °C. Fruit were removed from cool storage and ripened at room temperature using ethephon. The length of storage life was determined by the change in peel color to yellow, after this ethephon treatment. Fruit treated with 1-MCP + MAP had a storage life of 100 days. The storage life of control fruit (no 1-MCP and no MAP) was 20 days. Fruit held in PE bags without 1-MCP treatment had a 40 day storage life, and the same was found in fruit treated with 1-MCP but without PE bags. 1-MCP is an inhibitor of ethylene action, but also inhibited ethylene production, mainly through inhibition of ACC oxidase activity in the peel. MAP inhibited ethylene production mainly through inhibition of ACC oxidase, both in the peel and pulp. The combination of 1-MCP treatment and MAP storage resulted in much lower ethylene production due to inhibition of both ACC synthase and ACC oxidase activity.  相似文献   

7.
为明确一年生草本盐生植物费尔干猪毛菜(Salsola ferganica Drob.)病程相关蛋白基因SfPR1a (GenBank登录号为JQ670917)是否参与了植物对逆境胁迫的响应,采用qRT-PCR检测了该基因在不同组织部位和脱落酸(ABA)、茉莉酸(JAs)、乙烯合成直接前体(ACC)等相关激素胁迫及NaCl处理下的表达规律,同时对转基因烟草在盐、旱及丁香假单胞菌等胁迫下的抗性进行了鉴定。结果显示, SfPR1a基因在费尔干猪毛菜根中的表达量显著高于茎叶中,且受到ABA、JAs、ACC、NaCl的积极诱导;干旱胁迫下,转基因烟草的丙二醛(MDA)含量显著低于野生型烟草,显示出较强的抗旱表型;盐胁迫下,异源表达SfPR1a的转基因烟草幼苗生长显著优于野生型烟草;丁香假单胞菌攻毒后的转基因烟草叶片呈现严重的坏死反应,但植株的整体抗性表型显著优于野生型烟草;亚细胞定位结果显示该蛋白定位于植物细胞质外体空间。以上结果表明,费尔干猪毛菜病程相关蛋白SfPR1a基因参与了植物对非生物及生物胁迫的抗性。  相似文献   

8.
香蕉是重要的热带、亚热带地区作物,典型的呼吸跃变型果实,不耐贮藏.传统的保鲜方法存在诸多弊端,利用现代生物技术培育耐贮藏新品种,已经成为我们解决香蕉保鲜问题的重要课题.本研究利用果实特异性ACC合成酶基因反义植物表达载体,采用基因枪和农杆菌介导法转化海南主栽的巴西香蕉及红香蕉,并比较了不同转化方法及不同栽培品种的转化效果.结果表明农杆菌侵染法好于基因枪法,在农杆菌侵染方法中,对红香蕉的转化效果好于巴西香蕉.经PCR检测获得了9株转化植株,进一步采用Southern blot分析的方法,其中4株杂交信号较强,确认ACC合成酶反义基因已经整合进香蕉基因组中.本研究为香蕉的转基因研究和耐储藏新品种的培育奠定了一定的基础.  相似文献   

9.
Elongation growth is a typical feature of deepwater rice plants in response to submergence. The growth phenomenon is known to be induced by hypoxia, which results in the expression of genes implicated in ethylene biosynthesis. Ethylene is considered to trigger the growth response as it accumulates in the submerged tissues and submergence enhances the expression of 1-aminocyclopropane-1-carboxylate (ACC) synthase. However, ACC concentration in rice plants increases much faster after submergence than the activity of the ACC synthase genes studied previously. Here, we studied the expression characteristics of the fifth member of this gene family, OS-ACS5, and show that submergence induces the messenger concentration of this gene before the accumulation of ACC could be observed. OS-ACS5 may play a fundamental role in the growth-promoting increase in ethylene biosynthesis during the first hours of submergence in deepwater rice. This revised version was published online in August 2006 with corrections to the Cover Date.  相似文献   

10.
转codA基因提高番茄植株的耐热性   总被引：3，自引：0，他引：3  
以野生型番茄(cv. Moneymaker)和转codA番茄为材料,用不同温度(25、30、35、40、45和50℃)分别处理2 h,测定叶片净光合速率(Pn)、PSII最大光化学效率(Fv/Fm)、过氧化氢(H2O2)含量、丙二醛(MDA)含量、相对电导率(REC)和抗氧化酶活性等生理指标;42℃高温处理0、3和6 h后,检测热响应基因的表达量以及D1蛋白的含量,研究高温胁迫对上述参数的影响,探讨转codA基因提高番茄叶片耐热性的机制。。结果表明,高温胁迫下,转codA基因番茄叶片Pn和Fv/Fm的抑制程度明显低于野生型,H2O2、MDA的积累量以及REC均低于野生型,而且明显增强了过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、过氧化物酶(POD)和抗坏血酸过氧化物酶(APX)的活性。此外,转codA基因番茄叶片中抗氧化酶基因和热胁迫基因的表达水平均高于野生型,而D1蛋白的降解水平低于野生型。转codA基因番茄体内合成的甜菜碱提高了转基因番茄的耐热性,这与提高和维持较高的抗氧化酶活性、促进热激响应基因的表达及减缓D1蛋白的降解等有关。  相似文献   

11.
反义外壳蛋白基因介导的抗SCMV转基因玉米研究   总被引：6，自引：1，他引：6  
玉米矮花叶病（MDM）是一种世界性病害，在我国主要由甘蔗花叶病毒(SCMV)所致。为探索一条高效、安全的抗SCMV转基因途径，构建了无标记基因的SCMV反义外壳蛋白基因（cp）表达载体pACP。通过冻融法将该载体与抗除草剂标记基因(bar)载体分别导入农杆菌LBA4404,然后共转化玉米自交系综3的幼胚。通过除草剂梯度筛选，从抗性愈伤组织分化获得了35株再生苗。PCR检测证明，其中26株带有抗除草剂标记基因（bar），14株带有SCMV反义cp基因。这14株带有目的基因的玉米植株自交，其种子在田间种植成株行（T1代）。玉米T1代幼苗人工接种SCMV，筛选出2个抗病株率高于70%的株行。ELISA检测表明，抗病株SCMV含量极低，抗性达高抗水平。PCR检测表明，抗病性是反义cp基因作用的结果，并且获得了2株cp基因阳性而标记基因阴性的抗病株。  相似文献   

12.
Premature softening and tissue senescence occur in kiwifruit infected with Botrytis cinerea. While ethylene production is enhanced in infected fruit and B. cinerea produces ethylene on defined media in vitro the source of ethylene in this pathosystem is unclear. Ethylene production by B. cinerea was enhanced when methionine or ∝-keto-methylthiobutyric acid (KMBA) was added to a defined (modified Pratts) medium. Although 1-aminocyclopropane-1-carboxylic acid (ACC) did not stimulate ethylene production, ∝-aminooxyacetic acid (AOA) was inhibitory suggesting a role for a pyridoxal phosphate mediated enzyme reaction down stream from the methionine/KMBA stimulated ethylene biosynthetic pathway. Cobalt chloride (Co2+) was inhibitory, but after a 4-d lag period ethylene production from B. cinerea cultures containing methionine and Co2+ reached the same level as those without Co2+. [U 14C] methionine was converted to 14C-ethylene with high efficiency indicating that it is a direct precursor, while [2,3 14C]-ACC did not yield radioactively labelled ethylene. These results suggest that the ethylene biosynthetic pathway in B. cinerea does not involve ACC as a precursor and that the enzyme responsible for synthesising ethylene is similar to, but different from, ACC oxidase from higher plants. The ethylene biosynthetic pathway in B. cinerea is yet to be determined.  相似文献   

13.
乙烯与水稻细胞质雄性不育的关系   总被引：11，自引：1，他引：10  
田长恩  梁承邺 《作物学报》1999,25(1):116-119
从幼穗发育的IV到VII期,水稻细胞质雄性不育系(珍汕97A)幼穗和叶片的ACC含量和乙烯释放速率均高于其保持系(珍汕97B)。外施乙烯释放剂乙烯利使保持系花粉可育度明显下降;外施ACC合成酶抑制剂AVG引起两系幼穗ACC含量和乙烯释放速率下降,并使不育系花粉育性得以部分恢复,而在外施AVG的同时再施以乙烯利则AVG的恢复作用消失。  相似文献   

14.
    
Summary Simultaneous expression of a tobacco class I chitinase and a class I -1,3-glucanase gene in tomato resulted in increased fungal resistance, whereas transgenic tomato plants expressing either one of these genes were not protected against fungal infection. After infection with Fusarium oxysporum f.sp. lycopersici, a 36% to 58% reduction in disease severity was observed in resistant tomato lines. Two transgenic lines largely recovered from the initial infection by the time wild-type tomato plants had died.The overall results are consistent with the observation that class I chitinases and class I -1,3-glucanases synergistically inhibit the growth of fungi in vitro and provide the first experimental support to the hypothesis that such synergy can contribute to enhanced fungal resistance in planta.  相似文献   

15.
Transgenic Korean rice plants containing the cry1Ab gene were developed for resistance against yellow stem borer (Scirpophaga incertulas, YSB). More than 100 independent transgenic lines from three Korean varieties (P-I, P-II and P-III) were generated. The amount of Cry1Ab in transgenic T0 plants was as high as 2.88% of total soluble proteins. These levels were sufficient to cause 100% mortality of YSB larvae. The majority of T1 transgenic lines originated from the varieties P-I and P-II followed a Mendelian fashion of segregation. Deviation from the expected segregation ratio was observed in a small number of the transgenic lines of P-I and P-II origins. However, this deviation was primarily observed in the P-III originated lines. Segregation analysis of the T1 generation indicated that 1–3 copies of the cry1Ab gene were integrated into the genome of the majority of the transgenic lines originating from varieties P-I and P-II. Stunted and semi-fertile mutants were observed in some transgenic lines. These aberrations were either independent or closely linked to the introduced cry1Ab gene loci in different transgenic lines. Reduction in GUS expression levels and loss of toxicity against YSB larvae were found in some transgenic lines. The transgenic T3 and T4 lines causing 100% mortality of third instar YSB larvae in the lab were completely protected in the field. Analysis of important yield components on nine selected transgenic lines indicated that stem length, panicle length, grain number per panicle, and seed setting rates were reduced in transgenic plants compared to those in non-transgenic parental rice lines. Number of panicles per cluster, however, was significantly higher in transgenic plants. The numerical value of the average yield was in general greater in the controls than in all the transgenic lines, indicating some ‘yield drag’. Since some selected lines were highly resistant to the YSB with good yielding potential, they offer effective potential for use in insect resistance management programs.  相似文献   

16.
分析27个代表番茄不同发育阶段和生物反应的组织特异性、含有152 635个独立EST数据库的数码表达,发现果胶裂解酶基因 (pectate lyase, SlPEL) 和番茄AP2 Like (SlAPL)的转录受果实成熟的调节。以授粉后不同发育时期的番茄(品种为美味樱桃)果实为试材, 用半定量PCR和荧光实时定量PCR分析SlPEL的表达模式,结果表明,授粉后12 d,其表达水平明显上升;授粉后16~18 d,达到第一个小高峰;28 d到最高峰;从28 d到完全成熟逐步下降到第一个小高峰的水平。SlAPL的表达模式与SlPEL类似,但其表达启动的时期迟于SlPEL。从授粉后25 d,SlAPL转录启动;授粉后28~32 d,其转录水平上升到第一个小高峰;39 d达到最高峰,以后到完全成熟略有下降。该研究也印证利用EST的数据库进行基因数码表达分析的可行性。  相似文献   

17.
利用农杆菌介导法将反义Wx基因导入水稻的研究   总被引：3，自引：0，他引：3  
利用根癌农杆菌将含多基因(hpt选择标记基因、反义Wx基因、GFP和GUS报告基因)的pCAMBIA1304载体导入水稻品种(501R、中花9号和日本晴)的幼胚愈伤组织,分别在含35mg/L、45mg/L和65mg/L潮霉素浓度的筛选培养基上筛选获得抗性愈伤。后经PCR检测,从415株T0代再生植株中选出92株(其中501R、中花9号和日本晴分别为4株、43株和45株)。对这些转基因后代植株进行Southern blotting分析表明：hpt、反义Wx基因已经整合进植物基因组中。绝大多数转基因植株后代的表型正常。成熟种子直链淀粉含量分析表明,部分转基因植株的T1代种子中的直链淀粉含量有不同程度的下降,最低的已下降至6．3％,与对照相比下降了9．8％。  相似文献   

18.
小麦Mlo反义基因的转化及转基因植株的白粉病抗性分析   总被引：1，自引：0，他引：1  
采用基因枪法将小麦反义Mlo基因导入扬麦158和济麦20的幼胚愈伤组织中,在含除草剂的分化培养基上经两轮筛选,获得抗性再生植株。PCR检测、PCR-Southern杂交、基因组DNA斑点杂交和除草剂BASTA抗性分析结果证实已获得转基因扬麦158和济麦20阳性植株,荧光定量表达分析亦证明Mlo基因发生沉默。对T0和T1代转基因植株的白粉病抗性鉴定表明,有6个转基因株系高抗白粉病。对T1代转基因小麦接种白粉菌后孢子发育的显微观察结果显示,Mlo反义基因的导入明显加快了乳突的形成和维持时间,有效抑制了吸器的发育,因而使转Mlo反义基因材料表现抗病性。  相似文献   

19.
Summary For the past 10 years, the Andean-type Phaseolus vulgaris cultivar Kranskop has played an important role in South African bean production and breeding. Kranskop shares an ancestor with the internationally important Andean-type rust differential cultivar Redlands Pioneer. The Ur-13 gene in Kranskop and Redlands Pioneer gives protection to numerous internationally reported races of Uromyces appendiculatus and it is imperative to retain this gene in local breeding programmes.In this study, three co-dominant SCAR markers (SEAACMACC430/405, SEACAMCTT310/288 and SEAAGMCGT436 Hha I186/250) were used to trace the origin of Ur-13, and its presence in 71 germplasm accessions, including the international rust differential lines and additional sources of already characterized genes, as well as 78 breeding lines.Each marker was present in approximately 30% of the accessions tested. Only accessions belonging to the Kranskop or Redlands groups contained all three markers. Contrary to expectations, the first two markers, as well as Ur-13, appeared to have originated from a Middle-American-type parent of the Redlands group, California Small White 643, whereas the third marker probably came from the Andean-type cultivar Brown Beauty. This has important implications for the new set of differential lines, as Redlands Pioneer can no longer be regarded as a representative of the Andean gene pool. The markers, in particular SEAACMACC430/405, will be useful in tracing Ur-13 in large seeded breeding material, except where lines such as Mexico 309, PI 181996 and A 286 are used as donors of additional rust resistance genes, as these have the positive alleles of both SEAACMACC430/405 and SEACAMCTT310/288.Part of a Ph.D thesis submitted by the first author to the Department of Plant Sciences, University of the Free State.  相似文献   

20.
利用根癌农杆菌将含多基因(hyg选择标记基因、反义Wx 基因、PTA基因、GFP和GUS报告基因)的pCAMBIA1304载体导入水稻品种(501R、中花9号和日本晴)的幼胚愈伤组织,分别在含35mg/L、45mg/L和65mg/L潮霉素浓度的筛选培养基上筛选获得抗性愈伤。后经PCR检测,从415株T0代再生植株中选出92株(其中501R、中花9号和日本晴分别为4株、43株和45株)。对这些转基因后代植株进行Southern blotting分析表明：hpt、反义Wx 和 PTA基因已经整合进植物基因组中,绝大多数转基因植株后代的表型正常。成熟种子直链淀粉含量分析表明,部分转基因植株的T1代种子中的直链淀粉含量有部分程度的下降,最低的已下降至6.3%,与对照相比下降了9.8%。  相似文献   

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