Dixit Sameer, Upadhyay Santosh Kumar, Singh Harpal, Sidhu Om Prakash, Verma Praveen Chandra, K Chandrashekar
CSIR-National Botanical Research Institute, Council of Scientific and Industrial Research, Lucknow, Uttar Pradesh, India ; Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, 2-Rafi Marg, New Delhi, India.
PLoS One. 2013 Nov 5;8(11):e79664. doi: 10.1371/journal.pone.0079664. eCollection 2013.
Plants naturally emit methanol as volatile organic compound. Methanol is toxic to insect pests; but the quantity produced by most of the plants is not enough to protect them against invading insect pests. In the present study, we demonstrated that the over-expression of pectin methylesterase, derived from Arabidopsis thaliana and Aspergillus niger, in transgenic tobacco plants enhances methanol production and resistance to polyphagous insect pests. Methanol content in the leaves of transgenic plants was measured using proton nuclear spectroscopy (1H NMR) and spectra showed up to 16 fold higher methanol as compared to control wild type (WT) plants. A maximum of 100 and 85% mortality in chewing insects Helicoverpa armigera and Spodoptera litura larvae was observed, respectively when fed on transgenic plants leaves. The surviving larvae showed less feeding, severe growth retardation and could not develop into pupae. In-planta bioassay on transgenic lines showed up to 99 and 75% reduction in the population multiplication of plant sap sucking pests Myzus persicae (aphid) and Bemisia tabaci (whitefly), respectively. Most of the phenotypic characters of transgenic plants were similar to WT plants. Confocal microscopy showed no deformities in cellular integrity, structure and density of stomata and trichomes of transgenic plants compared to WT. Pollen germination and tube formation was also not affected in transgenic plants. Cell wall enzyme transcript levels were comparable with WT. This study demonstrated for the first time that methanol emission can be utilized for imparting broad range insect resistance in plants.
植物会自然释放甲醇这种挥发性有机化合物。甲醇对害虫有毒;但大多数植物产生的甲醇量不足以保护它们抵御害虫入侵。在本研究中,我们证明了拟南芥和黑曲霉来源的果胶甲酯酶在转基因烟草植物中的过表达可提高甲醇产量并增强对多食性害虫的抗性。使用质子核磁共振波谱(1H NMR)测量转基因植物叶片中的甲醇含量,光谱显示其甲醇含量比对照野生型(WT)植物高出多达16倍。以转基因植物叶片为食时,咀嚼式昆虫棉铃虫和斜纹夜蛾幼虫的死亡率分别最高达到100%和85%。存活的幼虫取食减少,生长严重受阻,无法发育成蛹。对转基因株系进行的植物体内生物测定表明,吸食植物汁液的害虫桃蚜(蚜虫)和烟粉虱(粉虱)的种群繁殖分别减少了多达99%和75%。转基因植物的大多数表型特征与野生型植物相似。共聚焦显微镜显示,与野生型相比,转基因植物的细胞完整性、气孔和毛状体的结构及密度均无畸形。转基因植物的花粉萌发和花粉管形成也未受影响。细胞壁酶转录水平与野生型相当。本研究首次证明甲醇排放可用于赋予植物广泛的抗虫性。