Overexpression of ghr-miR166b generates resistance against Bemisia tabaci infestation in Gossypium hirsutum plants
Overexpression of ghr-miR166b generates resistance against Bemisia tabaci infestation in Gossypium hirsutum plants
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DOI：
10.1007/s00425-018-2852-7
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发表时间：
2018-05-01
期刊：
影响因子：
4.3
通讯作者：
Khan, Jawaid A.
中科院分区：
文献类型：
--
作者：
Wamiq, Gazal;Khan, Jawaid A.
通过计算机模拟鉴定的陆地棉ghr-miR 166 b在烟粉虱线粒体ATP合酶中显示出多相容性靶标。其在植物中的过表达具有作为生物杀虫剂在减少B方面的潜力。烟粉虱（B.烟粉虱是半翅目昆虫，作为传播植物病毒的媒介，对植物造成巨大损失。在本研究中，G.靶向B的表达序列标签（EST）的多毛物编码的miRNA。基于序列互补性和miRNA-靶mRNA热力学，在计算机上鉴定了烟粉虱。在108 G中。hirsutum miRNAs，55靶向编码蛋白质的EST。其中，ghr-miR 166 b由于其对ATP合酶的固有亲和力而被选择。在G.陆地棉通过农杆菌介导的转化获得HS 6植株。共7个独立转化的（T-0）G.获得了陆地棉品系。观察到转基因株系中ghr-MIR 166 b的转录水平比非转化植物高2.0- 17倍。从转基因植物中分离的小RNA的Northern印迹分析证实了ghr-miR 166 b的存在。在饲喂具有最高水平的ghr-miR 166 B b表达的转基因系（HS 6 -166-30）的叶后，B.与未转化的叶相比，烟粉虱种群减少高达91%。此外，在全植物测定中，最高78%的B。在同一品系中观察到烟粉虱死亡率，而在B中则有增加。烟粉虱种群的非转化植物。我们的研究结果表明，ghr-miR 166 B可能靶向B的ATP合酶基因。烟粉虱，并随后在植物中的过表达，有可能作为生物农药减少B。烟粉虱种群和随之而来的烟粉虱传播病毒的传播。
In silico identified Gossypium hirsutum ghr-miR166b shows multi-compatible targets in mitochondrial ATP synthase of Bemisia tabaci. Its overexpression in planta has the potential to act as a biopesticide in reducing B. tabaci population, and consequently the spread of whitefly-transmitted plant viruses.Whiteflies (B. tabaci) are hemipterous insects that act as a vector to transmit plant viruses causing enormous losses to the plants. In the present study, G. hirsutum-encoded miRNAs targeting expressed sequence tags (ESTs) of B. tabaci, based on sequence complimentarity and miRNA-target mRNA thermodynamics, were in silico identified. Out of 108 G. hirsutum miRNAs, 55 targeted the protein encoding ESTs. Among them, ghr-miR166b was selected owing to its intrinsic affinity for ATP synthase. Its functional role was validated following expression of ghr-MIR166b (precursor) sequence in G. hirsutum cv. HS6 plants through Agrobacterium-mediated transformation. Total of seven independent transformed (T-0) G. hirsutum lines were obtained. The transcript level of ghr-MIR166b in the transgenic lines was observed to be 2.0- to 17-fold higher as compared to non-transformed plants. Northern-blot analysis of small RNAs isolated from the transgenic plants confirmed the presence of the ghr-miR166b. After feeding on the leaves of transgenic line (HS6-166-30) having highest level of ghr-miR166b expression, B. tabaci population was reduced up to 91% as compared to non-transformed leaves. Further, in the whole plant assay, a maximum of 78% B. tabaci mortality was observed in the same line, while there was an increase in B. tabaci population on the non-transformed plants. Our results revealed that ghr-miR166b supposedly targeting ATP synthase gene of B. tabaci, and subsequently its overexpression in planta has potential to act as biopesticide for reducing B. tabaci population and consequently spread of whitefly transmitted viruses.