在家蚕卵巢中对抗家蚕潜伏病毒的小RNA产生潜力,Archives of Insect Biochemistry and Physiology - X-MOL

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在家蚕卵巢中对抗家蚕潜伏病毒的小RNA产生潜力
Archives of Insect Biochemistry and Physiology ( IF 1.5 ) Pub Date : 2020-11-22 , DOI: 10.1002/arch.21761
Susumu Katsuma ₁ , Keisuke Shoji ₂ , Yutaka Suzuki ₃ , Masashi Iwanaga ₄

Affiliation

家蚕潜伏病毒（BmLV）是一种阳性单链昆虫RNA病毒，与植物黄斑病毒密切相关。BmLV首先是从家蚕卵巢细胞系BmN-4中分离出来的，该病毒已经感染了大多数家蚕细胞系。我们以前曾报道过小干扰RNA（siRNA）和PIWI相互作用RNA（piRNA）通路协同起作用，以维持正常BmN-4细胞生长的BmLV RNA量。另一方面，BmLV在家蚕双歧杆菌幼虫中不繁殖。在这里，我们将BmLV注射到了B. mori的幼虫体腔中，并研究了在siRNA和piRNA途径均活跃的幼虫卵巢中BmLV的积累，以研究这种体内耐药性是否受小RNA途径支配。与B. mori培养的细胞相比，BmLV注射的幼虫卵巢中RNA依赖性RNA聚合酶，外壳蛋白和p15基因的表达水平极低，这表明B. mori幼虫卵巢比B. mori更具抗性培养的细胞。我们还对从BmLV注射的幼虫卵巢制备的小RNA进行了测序，并将其定位到BmLV基因组中。尽管它们的量很小，但我们能够在卵巢中检测到BmLV衍生的小RNA。根据它们的长度分布和核苷酸偏倚，它们很可能是siRNA和piRNA。这些结果表明，家蚕双歧杆菌卵巢可能产生抗BmLV的小RNA，但是幼虫卵巢对BmLV的抗性并不依赖于RNA沉默途径。

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Potential for small RNA production against Bombyx mori latent virus in Bombyx mori ovaries

Bombyx mori latent virus (BmLV) is a positive, single‐stranded insect RNA virus closely related to plant maculaviruses. BmLV was first isolated from Bombyx mori ovary‐derived cell line BmN‐4, and this virus has already infected most B. mori‐derived cultured cell lines. We previously reported that small interfering RNA (siRNA) and PIWI‐interacting RNA (piRNA) pathways function cooperatively to maintain the amount of BmLV RNA for normal BmN‐4 cell growth. On the other hand, BmLV does not propagate in B. mori larvae. Here we conducted BmLV injection into the larval body cavities of B. mori, and examined BmLV accumulation in larval ovaries where siRNA and piRNA pathways are both active, to investigate whether this in vivo resistance is governed by small RNA pathways. Expression levels of RNA‐dependent RNA polymerase, coat protein, and p15 genes in BmLV‐injected larval ovaries were extremely low compared with those in B. mori cultured cells, indicating that B. mori larval ovaries are more resistant to BmLV than B. mori cultured cells. We also sequenced small RNAs prepared from BmLV‐injected larval ovaries and mapped them onto the BmLV genome. Although their amounts were very small, we were able to detect BmLV‐derived small RNAs in the ovaries. According to their length distribution and nucleotide bias, they were likely to be siRNAs and piRNAs. These results suggest that B. mori ovaries can potentially produce small RNAs against BmLV, but the resistance of larval ovaries against BmLV is not dependent on RNA silencing pathways.

更新日期：2020-11-22

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