Expression of endochitinase from Trichoderma harzianum in transgenic apple increases resistance to apple scab and reduces vigor
Expression of endochitinase from Trichoderma harzianum in transgenic apple increases resistance to apple scab and reduces vigor
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DOI：
10.1094/phyto.2000.90.1.72
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发表时间：
2000-01-01
期刊：
影响因子：
3.2
通讯作者：
Aldwinckle, HS
中科院分区：
文献类型：
--
作者：
Bolar, JP;Norelli, JL;Aldwinckle, HS
本研究的目的是通过转化生防菌哈茨木霉几丁质降解酶基因来提高苹果对黑星病的抗性。将几丁质内切酶基因以cDNA和基因组克隆的形式转入苹果cv.马歇尔麦金托什通过农杆菌转化。通过NPTII酶联免疫吸附试验和聚合酶链反应鉴定了15个品系为转基因品系，并通过Southern分析确认。通过酶法和Western分析，检测到不同品系间内切几丁质酶活性存在显著差异。用苹果黑星病菌接种8个品系的嫁接和自根植株。这些转基因株系表达内切几丁质酶比非转化的品种更耐。马歇尔麦金托什。与CV相比，疾病严重度。在测试的转基因品系中，马歇尔麦金托什减少了0至99.7%（病斑数）、0至90%（感染的叶面积百分比）和1至56%（回收的分生孢子）。几丁质内切酶对接种和未接种植株的生长也有负面影响。内切几丁质酶的产生水平与病害发生量和植株生长量呈显著负相关。
The goal of this research was to improve scab resistance of apple by transformation with genes encoding chitinolytic enzymes from the biocontrol organism Trichoderma harzianum. The endochitinase gene, as cDNA and genomic clones, was transferred into apple cv. Marshall McIntosh by Agrobacterium-transformation. A total of 15 lines were identified as transgenic by NPTII enzyme-linked immunosorbent assay and polymerase chain reaction and confirmed by Southern analysis. Substantial differences in endochitinase activity were detected among different lines by enzymatic assay and western analysis. Eight lines propagated as grafted and own-rooted plants were inoculated with Venturia inaequalis. Six of these transgenic lines expressing endochitinase were more resistant than nontransformed cv. Marshall McIntosh. Disease severity compared with cv. Marshall McIntosh was reduced by 0 to 99.7% (number of lesions), 0 to 90% (percentage of leaf area infected), and 1 to 56% (conidia recovered) in the transgenic lines tested. Endochitinase also had negative effects on the growth of both inoculated and uninoculated plants. There was a significant negative correlation between the level of endochitinase production and both the amount of disease and plant growth.