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      The role of ABA in triggering ethylene biosynthesis and ripening of tomato fruit

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          Abstract

          In order to understand more details about the role of abscisic acid (ABA) in fruit ripening and senescence of tomato, two cDNAs ( LeNCED1 and LeNCED2) which encode 9- cis-epoxycarotenoid dioxygenase (NCED) as a key enzyme in ABA biosynthesis, two cDNAs ( LeACS2 and LeACS4) which encode 1-aminocyclopropane-1-carboxylic acid (ACC) synthase, and one cDNA ( LeACO1) which encodes ACC oxidase involved in ethylene biosynthesis were cloned from tomato fruit using a reverse transcription-PCR (RT-PCR) approach. The relationship between ABA and ethylene during ripening was also investigated. Among six sampling times in tomato fruits, the LeNCED1 gene was highly expressed only at the breaker stage when the ABA content becomes high. After this, the LeACS2, LeACS4, and LeACO1 genes were expressed with some delay. The change in pattern of ACO activity was in accordance with ethylene production reaching its peak at the pink stage. The maximum ABA content preceded ethylene production in both the seeds and the flesh. The peak value of ABA, ACC, and ACC oxidase activity, and ethylene production all started to increase earlier in seeds than in flesh tissues, although they occurred at different ripening stages. Exogenous ABA treatment increased the ABA content in both flesh and seed, inducing the expression of both ACS and ACO genes, and promoting ethylene synthesis and fruit ripening, while treatment with fluridone or nordihydroguaiaretic acid (NDGA) inhibited them, delaying fruit ripening and softening. Based on the results obtained in this study, it was concluded that LeNCED1 initiates ABA biosynthesis at the onset of fruit ripening, and might act as an original inducer, and ABA accumulation might play a key role in the regulation of ripeness and senescence of tomato fruit.

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          Ethylene biosynthesis and action in tomato: a model for climacteric fruit ripening.

          Elucidating the mechanisms involved in ripening of climacteric fruit and the role that ethylene plays in the process are key to understanding fruit production and quality. In this review, which is based largely on research in tomato, particular attention is paid to the role of specific isoforms of ACC synthase and ACC oxidase in controlling ethylene synthesis during the initiation and subsequent autocatalytic phase of ethylene production during ripening. Recent information on the structure and role of six different putative ethylene receptors in tomato is discussed, including evidence supporting the receptor inhibition model for ripening, possible differences in histidine kinase activity between receptors, and the importance of receptor LeETR4 in ripening. A number of ethylene-regulated ripening-related genes are discussed, including those involved in ethylene synthesis, fruit texture, and aroma volatile production, as well as experiments designed to elucidate the ethylene signalling pathway from receptor through intermediate components similar to those found in Arabidopsis, leading to transcription factors predicted to control the expression of ethylene-regulated genes.
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            MOLECULAR BIOLOGY OF FRUIT MATURATION AND RIPENING.

            The development and maturation of fruits has received considerable scientific scrutiny because of both the uniqueness of such processes to the biology of plants and the importance of fruit as a significant component of the human diet. Molecular and genetic analysis of fruit development, and especially ripening of fleshy fruits, has resulted in significant gains in knowledge over recent years. Great strides have been made in the areas of ethylene biosynthesis and response, cell wall metabolism, and environmental factors, such as light, that impact ripening. Discoveries made in Arabidopsis in terms of general mechanisms for signal transduction, in addition to specific mechanisms of carpel development, have assisted discovery in more traditional models such as tomato. This review attempts to coalesce recent findings in the areas of fruit development and ripening.
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              A modified hot borate method significantly enhances the yield of high-quality RNA from cotton (Gossypium hirsutum L.).

              The isolation of biologically active RNA from cotton (Gossypium hirsutum L.) is difficult due to interference by high levels of endogenous phenolics, polysaccharides, and secondary metabolites. A modified hot borate procedure was developed to combat these cellular constituents during tissue homogenization, resulting in the quantitative recovery of RNA suitable for hybridization analysis, in vitro translation, and cDNA synthesis. The efficacy of several hot borate buffer adjuvants for the qualitative and quantitative recovery of leaf RNA was monitored by absorbance spectra, gel electrophoresis, protein, and cDNA synthesis. Of the buffer adjuvants evaluated, polyvinylpyrrolidone-40 (PVP-40) exhibited the single, most significant impact on the yield and quality of RNA isolated from cotton leaves, although inclusion of deoxycholate and/or Nonident-40 (NP-40) further enhanced the quality of the RNA. The unsurpassed qualitative and quantitative recovery of total RNA from cotton by hot borate buffer at alkaline pH, supplemented with PVP-40, deoxycholate, and/or NP-40 had also proven satisfactory for other recalcitrant plant species as well as for especially difficult tissue types.
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                Author and article information

                Journal
                J Exp Bot
                jexbot
                exbotj
                Journal of Experimental Botany
                Oxford University Press
                0022-0957
                1460-2431
                April 2009
                26 February 2009
                26 February 2009
                : 60
                : 6
                : 1579-1588
                Affiliations
                [1 ]College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, PR China
                [2 ]Department of Chemistry, Fudan University, Shanghai 200433, PR China
                Author notes
                [* ]To whom correspondence should be addressed. E-mail: pleng@ 123456cau.edu.cn
                Article
                10.1093/jxb/erp026
                2671613
                19246595
                e7a4a3fe-274e-4da5-b7c6-42fa7f35ef90
                © 2009 The Author(s).

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)

                History
                : 21 October 2008
                : 21 January 2009
                : 22 January 2009
                Categories
                Research Papers

                Plant science & Botany
                nced gene,fruit ripening,leacs2,ethylene,leaco1,aba,tomato
                Plant science & Botany
                nced gene, fruit ripening, leacs2, ethylene, leaco1, aba, tomato

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