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1.
Excessive softening is the main factor limiting fruit shelf life and storage. Transgenic plants modified in the expression of cell wall modifying proteins have been used to investigate the role of particular activities in fruit softening during ripening, and in the manufacture of processed fruit products. Transgenic experiments show that polygalacturonase (PG) activity is largely responsible for pectin depolymerization and solubilization, but that PG-mediated pectin depolymerization requires pectin to be de-methyl-esterified by pectin methylesterase (PME), and that the PG -subunit protein plays a role in limiting pectin solubilization. Suppression of PG activity only slightly reduces fruit softening (but extends fruit shelf life), suppression of PME activity does not affect firmness during normal ripening, and suppression of -subunit protein accumulation increases softening. All these pectin-modifying proteins affect the integrity of the middle lamella, which controls cell-to-cell adhesion and thus influences fruit texture. Diminished accumulation of either PG or PME activity considerably increases the viscosity of tomato juice or paste, which is correlated with reduced polyuronide depolymerization during processing. In contrast, suppression of -galactosidase activity early in ripening significantly reduces fruit softening, suggesting that the removal of pectic galactan side-chains is an important factor in the cell wall changes leading to ripening-related firmness loss. Suppression or overexpression of endo-(1\to4)-d-glucanase activity has no detectable effect on fruit softening or the depolymerization of matrix glycans, and neither the substrate nor the function for this enzyme has been determined. The role of xyloglucan endotransglycosylase activity in softening is also obscure, and the activity responsible for xyloglucan depolymerization during ripening, a major contributor to softening, has not yet been identified. However, ripening-related expansin protein abundance is directly correlated with fruit softening and has additional indirect effects on pectin depolymerization, showing that this protein is intimately involved in the softening process. Transgenic work has shown that the cell wall changes leading to fruit softening and textural changes are complex, and involve the coordinated and interdependent activities of a range of cell wall-modifying proteins. It is suggested that the cell wall changes caused early in ripening by the activities of some enzymes, notably -galactosidase and ripening-related expansin, may restrict or control the activities of other ripening-related enzymes necessary for the fruit softening process.  相似文献
2.
cDNA clones encoding homologues of expansins, a class of cell wall proteins involved in cell wall modification, were isolated from various stages of growing and ripening fruit of tomato (Lycopersicon esculentum). cDNAs derived from five unique expansin genes were obtained, termed tomato Exp3 to Exp7, in addition to the previously described ripening-specific tomato Exp1 (Rose et al. (1997) Proc Natl Acad Sci USA 94: 5955–5960). Deduced amino acid sequences of tomato Exp1, Exp4 and Exp6 were highly related, whereas Exp3, Exp5 and Exp7 were more divergent. Each of the five expansin genes showed a different and characteristic pattern of mRNA expression. mRNA of Exp3 was present throughout fruit growth and ripening, with highest accumulation in green expanding and maturing fruit, and lower, declining levels during ripening. Exp4 mRNA was present only in green expanding fruit, whereas Exp5 mRNA was present in expanding fruit but had highest levels in full-size maturing green fruit and declined during the early stages of ripening. mRNAs from each of these genes were also detected in leaves, stems and flowers but not in roots. Exp6 and Exp7 mRNAs were present at much lower levels than mRNAs of the other expansin genes, and were detected only in expanding or mature green fruit. The results indicate the presence of a large and complex expansin gene family in tomato, and suggest that while the expression of several expansin genes may contribute to green fruit development, only Exp1 mRNA is present at high levels during fruit ripening.  相似文献
3.
Plants of tomato (Lycopersicon esculentum Mill. cv. T5) were transformed with an antisense endo-1,4--glucanase (cellulase, EC 3.2.1.4) Cel2 transgene under the control of the constitutive cauliflower mosaic virus 35S promoter in order to suppress mRNA accumulation of Cel2. In two independent transgenic lines, Cel2 mRNA abundance was reduced by >95% in ripe fruit pericarp and ca. 80% in fruit abscission zones relative to non-transgenic controls. In both transgenic lines the softening of antisense Cel2 fruit pericarp measured using stress-relaxation analysis was indistinguishable from control fruit. No differences in ethylene evolution were observed between fruit of control and antisense Cel2 genotypes. However, in fruit abscission zones the suppression of Cel2 mRNA accumulation caused a significant (P<0.001) increase in the force required to cause breakage of the abscission zone at 4 days post breaker, an increase of 27% in one transgenic line and of 46% in the other transgenic line. Thus the Cel2 gene product contributes to cell wall disassembly occurring in cell separation during fruit abscission, but its role, if any, in softening or textural changes occurring in fruit pericarp during ripening was not revealed by suppression of Cel2 gene expression.  相似文献
4.
The soft flesh and deciduous fruit of pepper (Capsicum spp.) originated from the wild C. frutescens BG 2816 accession is a complete dominant trait controlled by the S gene. We constructed an F2 population from a cross of BG 2816 (SS) and the bell-type C. annuum cultivar Maor (ss) and determined that S cosegregated with the tomato fruit-specific endo-polygalacturonase (PG) gene. The soft flesh and deciduous fruit phenotypes were observed together in all F2 individuals, indicating a pleiotropic effect of PG on the two traits. We mapped S to pepper chromosome 10 in the region corresponding to that in which PG was previously mapped in tomato. Northern, RT-PCR and western analyses and enzyme activity assays, collectively, indicated that PG is not detected in green, breaker or red fruits of Maor, nor in green fruits of BG 2816. Accumulation of PG mRNA and protein was detected in the fruits of BG 2816, and it increased during ripening from breaker to red stages. The sequence analysis of partial PG cDNA isolated from BG 2816 revealed high homology (87% identity) with the tomato PG. The resemblance of the soft flesh and deciduous fruit phenotypes to PG-associated phenotypes in other fruit crops, the complete linkage between Sand PG, and the greater expression of PG in the fruits of BG 2816 than in those of Maor, all strongly indicate that PG is a candidate gene for S.  相似文献
5.
Cell walls extracted from cv. Rutgers, 7711 (ripening inhibited), and nor (non-ripening) tomato ( Lycopersicon eseulentum Mill.) pericarp tissue at various stages of post-maturation development have been separated into four distinct fractions and their carbohydrate composition characterized. The amount of ionically-associated, chelator-soluble (CDTA, cyclohexanediaminetetraacetic acid) uronic acid in 'Rutgers' fruit cell walls remained constant during ripening, whereas the amount of residual pectin, which was extracted with cold alkali (Na2CO3) and was apparently covalently bound, decreased. These changes did not occur in rin and nor mutant fruit at a similar chronological age. The galactose content in pectic polysaccharide preparations extracted from tomato cell walls with CDTA and Na2,CO3, decreased by 65% during ripening. A similar but diminished decrease also occurred in rin and nor fruit. A non-cellulosic polysaccharide(s) was present in walls which resisted extraction with Na-acetate/CDTA, Na2CO3, and 4 M KOH. In 'Rutgers' fruit, the content of galactose in this polysaccharide(s) decreased 44% during ripening, whereas little or no significant change was observed in rin or nor mutant fruit.  相似文献
6.
无论在室温还是低温条件下,肥城佛桃(Prunus persica cv.Fei-Cheng)果实在衰老软化过程中超弱发光均有明显的高峰出现,其出现时间与乙烯释放速率峰值相一致。乙烯合成抑制剂2-氨基乙氧基乙烯甘氨酸(2-aminoethoxyvinylglycine,AVG)与乙烯利(ethrel,化学名称2-氯乙基膦酸)处理结果显示,果实超弱发光并不随乙烯释放速率的增减而定量变化,说明果实乙烯释放与超弱发光变化没有直接的因果关系,超弱发光高峰的出现可能是果实衰老软化过程中代谢活跃的反映。  相似文献
7.
一种植物细胞壁松驰蛋白:膨胀素   总被引:5,自引:0,他引:5  
在植物细胞的生长过程中 ,多糖和蛋白质分泌到细胞壁里层 ,并形成具有一定机械强度的网络 ,这个网络是能伸展的 ,除非细胞停止生长。在细胞的生长过程中 ,一种细胞壁蛋白—膨胀素首次被鉴定出来具有使细胞壁的多糖网络疏松的能力 ,从而使膨压驱动的细胞扩大。膨胀素由两个多基因家族即α -膨胀素和 β -膨胀素多基因家族编码 ,每种基因的表达具有部位和细胞类型的特异性 ,但最新的研究也表明拟南芥中的膨胀素可以分为三个亚家族。越来越多的膨胀素基因从各种植物中鉴定出来 ,系统分析显示它们可能从一个共同的祖先基因进化而来。膨胀素的作用机理研究的还不是很清楚 ,但因为它们具有特别的功能 ,因此展现出良好的工业化应用前景。  相似文献
8.
Hemicelluloses were extracted from isolated tomato ( Lycopersicon esculentum Mill. cv. Rutgers) pericarp cell wall material at 3 different stages of ripeness with 4 M and 8 M KOH. Little change in molecular weight or composition of 4 M KOH-extracted material was observed during ripening. However, the composition of 8 M KOH-extracted material changed, and a relative increase in polymers of < 40 kDa was observed during ripening. Changes in glycosyl linkage composition of the 8 M KOH hemicellulosic material were detected, including increases in 4-linked mannosyl, 4,6-linked mannosyl, and 4-linked glucosyl, and decreases in 5-linked arabinosyl residues in polymers of < 40 kDa, and decreases in terminal glocosyl residues in polymers of > 40 kDa. These data may indicate that de novo hemicellulose synthesis occurs throughout tomato fruit ripening, even at the red ripe stage.  相似文献
9.
以中华猕猴桃(ActinidiachinensisPlanch.)果实为试材,木葡聚糖内糖基转移酶(XET)cDNA为探针,研究果实成熟进程中XETmRNA的变化规律,探讨XET在果实后熟软化过程的作用。结果表明,20℃下外源乙烯处理可促进XETmRNA的积累,且这种效应因乙烯处理时间的加长而加强,进而加速了果实软化;0℃处理可抑制XETmRNA的增加,延缓果实软化,但当果实转入20℃后熟时,果实硬度迅速下降,而XETmRNA水平变化不明显。认为XET可能只是一种诱导酶,由它引起的细胞壁解聚并非是猕猴桃果实后熟软化的关键因子。  相似文献
10.
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