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Chapter 28 - RNA interference against RNA viruses in plants

Viruses are the biotrophic parasites of plants, cause a wide variety of symptoms, and are responsible for considerable yield loss in a variety of crops. The genera, viz., Tospovirus, Ilarvirus, Cucumovirus, and Potyvirus, are the major RNA plant viruses that infect cultivated crops worldwide which leads to huge loss. The virus disease management relies upon the use of prophylactic measures, cultural practices, planting resistant cultivars, vector control options, and molecular approaches. The efforts made through the conventional breeding program by developing resistant cultivars against viruses that are not successful in all crops due to unavailability of resistant sources. Alternatively, the RNA silencing technology has been explored as an advanced biotechnological tool for the control of virus diseases in crop plants. The antiviral mechanism through RNA silencing in plants is also known as RNA interference (RNAi) or posttranscriptional gene-silencing (PTGS). RNA silencing is a surveillance response induced by double-stranded RNA (dsRNA), wherein the dsRNA is cleaved into short small interfering RNA (siRNA), which is the key attribute for sequence-specific degradation of mRNA. The plants expressing sense and antisense gene fragments of viruses have shown resistance to virus. The inverted repeat constructs producing self-complementary RNAs (hairpin RNAs) from viral origin have efficiently induced virus resistance in transgenic plants through RNA-silencing mechanism. RNA silencing has expedited the molecular basis of plant antiviral systems, since it allows more accurate downregulation of gene expression without influencing the expression of other genes.

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Chapter 24 - Genetic analysis of banana bract mosaic virus and its management

Banana bract mosaic virus belonging to the genus potyvirus is one of the serious banana-infecting viruses causing bract mosaic disease which is responsible for a significant negative impact on the production and productivity in India and the Philippines. The virus is a flexuous rod-shaped containing single-stranded positive-sense RNA genome with a length of 9708 to 9713 nucleotides long and codes for a polyprotein of 3125 amino acids, and the polyprotein gets cleaved by nine proteases to yield 10 functional matured proteins. This virus induces mosaic symptoms on the leaf, leaf sheath, midrib, fruits, petiole, and malformation fingers and bunches. So far BBrMV is reported to cause diseases on banana, cardamom, and flowering ginger, and its distribution is limited to a few countries including India. As on date, only four full-length complete genome sequences are reported and nucleotide sequences of the complete genome of the BBrMV-TRY (banana) isolate were 96.7% identical with BBrMV-WAY (cardamom) followed by BBrMV-PHI (banana) (94.6%) and BBrMV-HAW (flowering ginger) (94.4%). The genetic diversity of BBrMV isolates was analyzed based on the sequences available in the public database. Based on the coat-protein-encoding gene of BBrMV isolates, two distinct isolates, TN14 and TN16, were identified which had a divergence of 19.8% and 17% at nt and 15.4% and 7.7% at aa level, respectively, with other isolates. The evolutionary distance, polymorphism at the sequence level, the presence of SNPs and INDELs, and the codon-based analysis and genetic differentiation between the BBrMV populations based on the sequence of three genes, viz. the HC-Pro, VPg, and CP were performed. Based on SNPs analysis, a maximum of 74, 36, 177 SNPs, and a minimum of 24, 17, 15SNPs, for HC-Pro, VPg, and CP genes were detected. The codon-based selection analysis of HC-Pro, VPg, and CP revealed that most of the codons fell under negative selection, and some were under positive selection. The gene flow between BBrMV populations was infrequent. Phylogenetic analysis and the results revealed the closeness of BBrMV-TRY with BBrMV cardamom isolate. Recombination analysis showed only four potential recombinants in both the HC-Pro and CP genes, respectively, whereas VPg had a single recombinant. In the management of this disease, the detection plays an important role in eliminating the virus from mother plants used for mass tissue culture propagation, confirming the elimination of the virus attempted by meristem tip culture and other therapies. The higher dose of fertilizer leads to the compensation of yield loss due to this virus in banana. Molecular approaches like transgenics, RNAi, and genome editing can be applied to develop resistance against this virus in banana, and so far only a few attempts were made to develop genetically engineered plants using these approaches.

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Chapter 9 - Cucurbit-infecting poleroviruses: Emerging plant RNA viruses in Indian agriculture

The genus Polerovirus is currently ascribed to the family Solemoviridae. Cucurbit aphid-borne yellows virus (CABYV) was the first cucurbit-infecting polerovirus recorded in the world from France in 1988 infecting cucumber, melon, and zucchini squash plants. Subsequently, this has been documented worldwide infecting various cucurbits within a span of 30 years from its first documentation, indicating its rapid global distribution, hence gaining the status of an “emerging virus.” Globally, eight Polerovirus species have been documented to cause economically important diseases in cucurbit crops. However, the occurrence of these viruses was not known in India until the first report of CABYV in 2017. In the recent past, the incidences of cucurbit-infecting poleroviruses have been increasing worldwide in different hosts and geographic locations causing 10%–100% crop losses in several crops. However, their impact on different crops has remained to be estimated in India. This book chapter documents the up-to-date information available on the occurrence and distribution, biology, molecular biology, detection and diagnosis, epidemiology, and management aspects of the cucurbit-infecting poleroviruses in terms of global and Indian contexts as well as the future thrust areas of research investigations.

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Chapter 13 - Virus-vector interaction and transmission in plants

Viruses are unique pathogens of agriculturally important crops. Transmission of plant viruses via insect vectors is an interesting mechanism in the life cycle of viruses and insects as well. Virus-vector interaction develops the understanding of the transmission mechanisms involved. Plant viruses spread horizontally and vertically through insects depending on the transmission mode. Plant viruses interact with their vectors in different ways for their survival and spread. Different proteins in viruses, hosts, and insects are involved in the transmission and acquisition of plant viruses from different sources, although insect proteins are involved in virus transmission but are poorly characterized. Hitherto, insect proteins involved in virus transmission have been poorly characterized. Identification of different proteins and their role involved in molecular mechanisms in virus acquisition and transmission through insects are the key areas in the research gap and recent advances. Among the other proteins, cuticular proteins have been emerged as the vital molecules in plant virus transmission. This chapter will describe the role of insects in the life cycles of viruses and hosts. The interaction of virus-vector-host will also be elaborated through the role of different proteins and molecular mechanisms in this chapter. These understanding and interactions and mechanisms will be helpful to formulate the management strategies for plant viruses in agricultural crops.

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