The MADS box gene, FOREVER YOUNG FLOWER, acts as a repressor controlling floral organ senescence and abscission in Arabidopsis

The ectopic expression of a MADS box gene FOREVER YOUNG FLOWER (FYF) caused a significant delay of senescence and a deficiency of abscission in flowers of transgenic Arabidopsis. The defect in floral abscission was found to be due to a deficiency in the timing of cell separation of the abscission zone cells. Down-regulation of INFLORESCENCE DEFICIENT IN ABSCISSION (IDA) may contribute to the delay of the floral abscission in 35S:FYF flowers. FYF was found to be highly expressed in young flowers prior to pollination and was significantly decreased after pollination, a pattern that correlated with its function. Ethylene insensitivity in senescence/abscission and the down-regulation of ETHYLENE RESPONSE DNA-BINDING FACTOR 1 (EDF1) and EDF2, downstream genes in the ethylene response, in 35S:FYF Arabidopsis suggested a role for FYF in regulating senescence/abscission by suppressing the ethylene response. This role was further supported by the fact that 35S:FYF enhanced the delay of flower senescence/abscission in ethylene response 1 (etr1), ethylene-insensitive 2 (ein2) and constitutive triple response 1 (ctr1) mutants, which have defects in upstream genes of the ethylene signaling pathway. The presence of a repressor domain in the C-terminus of FYF and the enhancement of the delay of senescence/abscission in FYF+SRDX (containing a suppression motif) transgenic plants suggested that FYF acts as a repressor. Indeed, in FYF-DR+VP16 transgenic dominant-negative mutant plants, in which FYF was converted to a potent activator by fusion to a VP16-AD motif, the senescence/abscission of the flower organs was significantly promoted, and the expression of BOP2, IDA and EDF1/2 was up-regulated. Our data suggest a role for FYF in controlling floral senescence/abscission by repressing ethylene responses and regulating the expression of BOP2 and IDA in Arabidopsis.     

Defence responses regulated by jasmonate and delayed senescence caused by ethylene receptor mutation contribute to the tolerance of petunia to Botrytis cinerea

Ethylene and jasmonate (JA) have powerful effects when plants are challenged by pathogens. The inducible promoter‐regulated expression of the Arabidopsis ethylene receptor mutant ethylene‐insensitive1‐1 (etr1‐1) causes ethylene insensitivity in petunia. To investigate the molecular mechanisms involved in transgenic petunia responses to Botrytis cinerea related to the ethylene and JA pathways, etr1‐1‐expressing petunia plants were inoculated with Botrytis cinerea. The induced expression of etr1‐1 by a chemical inducer dexamethasone resulted in retarded senescence and reduced disease symptoms on detached leaves and flowers or intact plants. The extent of decreased disease symptoms correlated positively with etr1‐1 expression. The JA pathway, independent of the ethylene pathway, activated petunia ethylene response factor (PhERF) expression and consequent defence‐related gene expression. These results demonstrate that ethylene induced by biotic stress influences senescence, and that JA in combination with delayed senescence by etr1‐1 expression alters tolerance to pathogens.     

The central role of PhEIN2 in ethylene responses throughout plant development in petunia

The plant hormone ethylene regulates many aspects of growth and development. Loss-of-function mutations in ETHYLENE INSENSITIVE2 (EIN2) result in ethylene insensitivity in Arabidopsis, indicating an essential role of EIN2 in ethylene signaling. However, little is known about the role of EIN2 in species other than Arabidopsis. To gain a better understanding of EIN2, a petunia (Petunia x hybrida cv Mitchell Diploid [MD]) homolog of the Arabidopsis EIN2 gene (PhEIN2) was isolated, and the role of PhEIN2 was analyzed in a wide range of plant responses to ethylene, many that do not occur in Arabidopsis. PhEIN2 mRNA was present at varying levels in tissues examined, and the PhEIN2 expression decreased after ethylene treatment in petals. These results indicate that expression of PhEIN2 mRNA is spatially and temporally regulated in petunia during plant development. Transgenic petunia plants with reduced PhEIN2 expression were compared to wild-type MD and ethylene-insensitive petunia plants expressing the Arabidopsis etr1-1 gene for several physiological processes. Both PhEIN2 and etr1-1 transgenic plants exhibited significant delays in flower senescence and fruit ripening, inhibited adventitious root and seedling root hair formation, premature death, and increased hypocotyl length in seedling ethylene response assays compared to MD. Moderate or strong levels of reduction in ethylene sensitivity were achieved with expression of both etr1-1 and PhEIN2 transgenes, as measured by downstream expression of PhEIL1. These results demonstrate that PhEIN2 mediates ethylene signals in a wide range of physiological processes and also indicate the central role of EIN2 in ethylene signal transduction.     

花衰老相关的乙烯信号转导基因研究进展

乙烯在许多切花衰老过程中起着重要的调节作用,不同的植物乙烯信号转导组分在花衰老过程中有不同的转录调节特性。根据乙烯信号转导标准模式,通过调节乙烯信号转导基因表达能够调控花对乙烯的敏感性,深入研究乙烯信号转导机制;可能有多条途径可延缓切花衰老。综述了香石竹和月季等几种观赏植物在花衰老过程中乙烯受体和乙烯信号转导基因表达及特性。     

Mutational analysis of the ethylene receptor ETR1. Role of the histidine kinase domain in dominant ethylene insensitivity

The ethylene receptor family of Arabidopsis consists of five members, one of these being ETR1. The N-terminal half of ETR1 contains a hydrophobic domain responsible for ethylene binding and membrane localization. The C-terminal half of the polypeptide contains domains with homology to histidine (His) kinases and response regulators, signaling motifs originally identified in bacteria. The role of the His kinase domain in ethylene signaling was examined in planta. For this purpose, site-directed mutations were introduced into the full-length wild-type ETR1 gene and into etr1-1, a mutant allele that confers dominant ethylene insensitivity on plants. The mutant forms of the receptor were expressed in Arabidopsis and the transgenic plants characterized for their ethylene responses. A mutation that eliminated His kinase activity did not affect the ability of etr1-1 to confer ethylene insensitivity. A truncated version of etr1-1 that lacks the His kinase domain also conferred ethylene insensitivity. Possible mechanisms by which a truncated version of etr1-1 could exert dominance are discussed.     

Hormonal and stress induction of the gene encoding common bean acetyl-coenzyme A carboxylase

Regulation of the cytosolic acetyl-coenzyme A carboxylase (ACCase) gene promoter from common bean (Phaseolus vulgaris) was studied in transgenic Arabidopsis (Arabidopsis thaliana) plants using a beta-glucuronidase (GUS) reporter gene fusion (PvACCase::GUS). Under normal growth conditions, GUS was expressed in hydathodes, stipules, trichome bases, flowers, pollen, and embryos. In roots, expression was observed in the tip, elongation zone, hypocotyl-root transition zone, and lateral root primordia. The PvACCase promoter was induced by wounding, Pseudomonas syringae infection, hydrogen peroxide, jasmonic acid (JA), ethylene, or auxin treatment. Analysis of PvACCase::GUS expression in JA and ethylene mutants (coronatine insensitive1-1 [coi1-1], ethylene resistant1-1 [etr1-1], coi1-1/etr1-1) suggests that neither JA nor ethylene perception participates in the activation of this gene in response to wounding, although each of these independent signaling pathways is sufficient for pathogen or hydrogen peroxide-induced PvACCase gene expression. We propose a model involving different pathways of PvACCase gene activation in response to stress.     

The influence of ethylene perception on sex expression in melon (<Emphasis Type="Italic">Cucumis melo </Emphasis>L.) as assessed by expression of the mutant ethylene receptor, <Emphasis Type="Italic">At-etr1-1</Emphasis>, under the control of constitutive and floral targeted promoters

Sexual diversity expressed by Curcurbitaceae species is a primary example of developmental plasticity in plants. Ethylene, which promotes femaleness (carpel development), plays a key role in sex determination. We sought to determine the critical location for ethylene perception in developing floral primodia. The dominant negative Arabidopsis ethylene response mutant gene, etr1-1, was introduced into melon (Cucumis melo L.) plants under control of the constitutive cauliflower mosaic virus (CaMV) 35S promoter, or floral-targeted Apetela3 (AP3) and Crab’s Claw (CRC) promoters, which in Arabidopsis, promote expression in petal and stamen, and carpel and nectary primordia, respectively. Based on effects of exogenous ethylene, it was predicted that inhibition of ethylene perception by carpel primordia would inhibit carpel development. Constitutive expression of etr1-1 caused several phenotypes associated with ethylene insensitivity, verifying that etr1-1 inhibits ethylene perception in the heterologous melon system. Carpel-bearing bud production was essentially abolished in 35S::etr1-1 melons, providing direct demonstration of the requirement for ethylene perception for carpel development. CRC::etr1-1 plants, however, showed enhanced femaleness as manifested by earlier and increased number of carpel-bearing buds, and production of female (rather than bisexual) buds. Despite increased carpel-bearing bud formation, a greater proportion of the CRC::etr1-1 carpel-bearing buds aborted before anthesis. AP3::etr1-1 plants showed increased maleness by nearly exclusive staminate flower production, and poorly developed carpels in the rare bisexual flowers. These results indicate that ethylene perception by the stamen (or petal) primordia plays a critical role in promoting carpel development at the time of sex determination, while ethylene perception by the carpel is important for maturation of carpel-bearing flowers to anthesis.     

The role of ethylene in the senescence of carnation flowers,a review

The senescence of flower petals is a highly regulated developmental process which requires active gene expression and protein synthesis. The biochemical changes associated with petal senescence in carnation flowers include an increase in hydrolytic enzymes, degradation of macro-molecules, increased respiratory activity and a climacteric-like increase in ethylene production. It is clear that the gaseous phytohormone ethylene plays a critical role in the regulation and coordination of senescence processes. Many reviews on physiology and mode of action of ethylene are available. Molecular cloning led to the isolation of genes involved in ethylene biosynthesis and action. This review describes the current status of the studies on regulation of ethylene biosynthesis and ethylene response in carnation flowers. An overview is given of studies on senescence-related gene expression and possibilities to improve postharvest longevity by genetic engineering.     

Role of cytokinins in carnation flower senescence

Stem and leaf tissues of carnation (Dianthus caryophyllus) plants appear to contain a natural antisenescence factor since removal of most of these tissues from cut carnation flowers hastened their senescence. However, kinetin (5-10 μg/ml) significantly delayed senescence of flowers with stem and leaf tissues removed. In addition, the life span of cut flowers with intact (30-cm) stems was increased with kinetin treatment. Peak ethylene production by presenescent flowers was reduced 55% or more with kinetin treatment and was delayed by 1 day. Kinetin-treated flowers were less responsive to applied ethylene (100 μl/l for 3 hours) than untreated flowers. Possible natural roles of cytokinins in carnation flower senescence are discussed.     

不同结构的外源ACO基因导入香石竹对瓶插寿命的影响

以香石竹叶片为外植体 ,利用根癌农杆菌 (Agrobacteriumtumefaciens)介导法 ,将香石竹ACC氧化酶 (ACO)基因核DNA的正义 (sense)、反义 (antisense)、正义重复 (sensedirectrepeat)和反义重复 (antisensedirectrepeat)等 4种T DNA结构导入香石竹‘Master’品种。经Southern杂交检测证明目的基因已整合到香石竹基因组 ,共获得 14个转化株系。在 25℃条件下比较瓶插寿命 ,对照植株花朵瓶插寿命为 5.8d ,多数转化株系花朵瓶插寿命达 11d ,最长者可达 12.8d。大多数转基因株系切花衰老过程中乙烯释放量显著减少 ,部分转基因株系切花衰老过程中几乎检测不到乙烯 ,而对照有明显的峰值。通过对本研究转化ACO基因核DNA与前人转化ACO基因cDNA延长瓶插寿命比较以及对不同T DNA结构的转化抑制内源基因表达的程度进行比较后 ,初步判断用核DNA转化后对内源基因的抑制效果与cDNA相当甚至更明显 ,反义基因可以比正义基因更有效地抑制内源的同源基因的表达 ,转重复基因比转单个基因能更有效地抑制内源的同源基因的表达。     

Members of the tomato LeEIL (EIN3-like) gene family are functionally redundant and regulate ethylene responses throughout plant development

The plant hormone ethylene regulates many aspects of growth, development and responses to the environment. The Arabidopsis ETHYLENE INSENSITIVE3 (EIN3) protein is a nuclear-localized component of the ethylene signal-transduction pathway with DNA-binding activity. Loss-of-function mutations in this protein result in ethylene insensitivity in Arabidopsis. To gain a better understanding of the ethylene signal-transduction pathway in tomato, we have identified three homologs of the Arabidopsis EIN3 gene (LeEILs). Each of these genes complemented the ein3-1 mutation in transgenic Arabidopsis, indicating that all are involved in ethylene signal transduction. Transgenic tomato plants with reduced expression of a single LeEIL gene did not exhibit significant changes in ethylene response; reduced expression of multiple tomato LeEIL genes was necessary to reduce ethylene sensitivity significantly. Reduced LeEIL expression affected all ethylene responses examined, including leaf epinasty, flower abscission, flower senescence and fruit ripening. Our results indicate that the LeEILs are functionally redundant and positive regulators of multiple ethylene responses throughout plant development.     

Role of the gynoecium in natural senescence of carnation (Dianthus caryophyllus L.) flowers

Although the role of the gynoecium in natural senescence of the carnation flower has long been suggested, it has remained a matter of dispute because petal senescence in the cut carnation flower was not delayed by the removal of gynoecium. In this study, the gynoecium was snapped off by hand, in contrast to previous investigations where removal was achieved by forceps or scissors. The removal of the gynoecium by hand prevented the onset of ethylene production and prolonged the vase life of the flower, demonstrating a decisive role of the gynoecium in controlling natural senescence of the carnation flower. Abscisic acid (ABA) and indole-3-acetic acid (IAA), which induced ethylene production and accelerated petal senescence in carnation flowers, did not stimulate ethylene production in the flowers with gynoecia removed (-Gyn flowers). Application of 1-aminocyclopropane-1-carboxylate (ACC), the ethylene precursor, induced substantial ethylene production and petal wilting in the flowers with gynoecia left intact, but was less effective at stimulating ethylene production in the -Gyn flowers and negligible petal in-rolling was observed. Exogenous ethylene induced autocatalytic production of the gas and petal wilting in the -Gyn flowers. These results indicated that ethylene generated in the gynoecium triggers the onset of ethylene production in the petals of carnation during natural senescence.     

The role of ethylene in the senescence of carnation flowers, a review

The senescence of flower petals is a highly regulated developmental process which requires active gene expression and protein synthesis. The biochemical changes associated with petal senescence in carnation flowers include an increase in hydrolytic enzymes, degradation of macro-molecules, increased respiratory activity and a climacteric-like increase in ethylene production. It is clear that the gaseous phytohormone ethylene plays a critical role in the regulation and coordination of senescence processes. Many reviews on physiology and mode of action of ethylene are available. Molecular cloning led to the isolation of genes involved in ethylene biosynthesis and action. This review describes the current status of the studies on regulation of ethylene biosynthesis and ethylene response in carnation flowers. An overview is given of studies on senescence-related gene expression and possibilities to improve postharvest longevity by genetic engineering.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AIB -amino-isobutyric acid - AOA amino oxyacetic acid - AVG aminoathoxyvinyl glycine - DACP diazocyclopentadiene - EFE ethylene forming enzyme - MACC malonyl 1-aminocyclopropane-1-carboxylic acid - MTA 5-methylthio-adenosine - NBD 2,5 norbornadiene - ppb parts per billion - SAM S-adenosyl-methionine - STS silver thiosulphate     

Ethylene regulates the timing of leaf senescence in Arabidopsis

The plant hormone ethylene influences many aspects of plant growth and development, including some specialized forms of programmed senescence such as fruit ripening and flower petal senescence. To study the relationship between ethylene and leaf senescence, etr1-1, an ethylene-insensitive mutant in Arabidopsis, was used. Comparative analysis of rosette leaf senescence between etr1-1 and wild-type plants revealed that etr1-1 leaves live approximately 30% longer than the wild-type leaves. Delayed leaf senescence in etr1-1 coincided with delayed induction of senescence-associated genes (SAGs) and higher expression levels of photosynthesis-associated genes (PAGs). In wild-type plants, exogenous ethylene was able to further accelerate induction of SAGs and decrease expression of PAGs. The extended period of leaf longevity in etr1-1 was associated with low levels of photosynthetic activity. Therefore, the leaves in etr1-1 functionally senesced even though the apparent life span of the leaf was prolonged.