Engineering melon plants with improved fruit shelf life using the TILLING approach.
Engineering melon plants with improved fruit shelf life using the TILLING approach.
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DOI：
10.1371/journal.pone.0015776
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发表时间：
2010-12-30
期刊：
影响因子：
3.7
通讯作者：
Bendahmane A
中科院分区：
文献类型：
--
作者：
Dahmani-Mardas F;Troadec C;Boualem A;Lévêque S;Alsadon AA;Aldoss AA;Dogimont C;Bendahmane A
水果的成熟和软化是影响食物供应、水果营养价值以及人类健康的关键特征。乙烯对更年期果实的成熟具有诱导作用，是控制果实过熟导致果实软化变质的主要目标之一。拟南芥和番茄乙烯途径的鉴定鉴定了控制果实成熟的关键基因。为了提高甜瓜的保质期，我们进行了一项转化研究实验。我们在一个雌雄同株和更年期的甜瓜品系上建立了TILLING平台，克隆了控制乙烯产生的基因，并筛选了导致果实延长保质期的诱导突变。鉴定了乙烯合成酶ACC氧化酶1的两个错义突变L124F和G194D，并对突变植株进行了果实成熟的特征分析。L124F突变是一种保守突变，发生在远离酶活性位点的地方，因此预计不会影响乙烯的产生，从而影响果实的成熟。相比之下，G194D的修饰发生在一个高度保守的氨基酸位置，通过晶体分析预测，会影响酶的活性。表型分析显示，G194D突变体果实成熟完全延迟，变黄，保质期延长，与预测的一样，L124F突变没有影响。我们构建了4023个甜瓜M2家族的突变体集合。基于11个基因的TILLING，我们计算了每573kb 1个突变的总突变率，并在每TILLING kobase中鉴定出8个等位基因。我们还发现了一个TILLING突变体，具有延长的水果保质期。这项工作证明了TILLING作为一种反向遗传工具来改良作物品种的有效性。由于葫芦是植物生物学不同领域的模式物种，我们期望开发的工具将被科学界广泛利用。
Fruit ripening and softening are key traits that have an effect on food supply, fruit nutritional value and consequently, human health. Since ethylene induces ripening of climacteric fruit, it is one of the main targets to control fruit over ripening that leads to fruit softening and deterioration. The characterization of the ethylene pathway in Arabidopsis and tomato identified key genes that control fruit ripening. To engineer melon fruit with improved shelf-life, we conducted a translational research experiment. We set up a TILLING platform in a monoecious and climacteric melon line, cloned genes that control ethylene production and screened for induced mutations that lead to fruits with enhanced shelf life. Two missense mutations, L124F and G194D, of the ethylene biosynthetic enzyme, ACC oxidase 1, were identified and the mutant plants were characterized with respect to fruit maturation. The L124F mutation is a conservative mutation occurring away from the enzyme active site and thus was predicted to not affect ethylene production and thus fruit ripening. In contrast, G194D modification occurs in a highly conserved amino acid position predicted, by crystallographic analysis, to affect the enzymatic activity. Phenotypic analysis of the G194D mutant fruit showed complete delayed ripening and yellowing with improved shelf life and, as predicted, the L124F mutation did not have an effect. We constructed a mutant collection of 4023 melon M2 families. Based on the TILLING of 11 genes, we calculated the overall mutation rate of one mutation every 573 kb and identified 8 alleles per tilled kilobase. We also identified a TILLING mutant with enhanced fruit shelf life. This work demonstrates the effectiveness of TILLING as a reverse genetics tool to improve crop species. As cucurbits are model species in different areas of plant biology, we anticipate that the developed tool will be widely exploited by the scientific community.
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影响因子：
56.9
作者：
Boualem, Adnane;Fergany, Mohamed;Bendahmane, Abdelhafid
通讯作者：
Bendahmane, Abdelhafid
影响因子：
56.9
作者：
Alonso, JM;Stepanova, AN;Ecker, JR
通讯作者：
Ecker, JR
影响因子：
7.2
作者：
Caldwell, DG;McCallum, N;Waugh, R
通讯作者：
Waugh, R
影响因子：
5.3
作者：
Deleu W;Esteras C;Roig C;González-To M;Fernández-Silva I;Gonzalez-Ibeas D;Blanca J;Aranda MA;Arús P;Nuez F;Monforte AJ;Picó MB;Garcia-Mas J
通讯作者：
Garcia-Mas J
影响因子：
5.4
作者：
Fernandez-Silva, I.;Eduardo, I.;Monforte, Antonio Jose
通讯作者：
Monforte, Antonio Jose