First report of Fusarium falciforme causing basal rot of onion (Allium cepa) in India.

Onion (Allium cepa, L) is a very important vegetable crop in India. India is the second largest producer of onion in the world and the crop is grown on more than 1.22 million hectares. Fusarium Basal Rot (FBR) is an economically important diseasef onion that causes considerable losses in onion production up to 50% in field and 30-40% during post-harvest storage of bulbs (Gupta and Gupta 2013; Rajamohan et al. 2019). Onion plants showing chlorosis, twisting, wilting, necrosis, bulb discoloration, rot in the basal parts of bulb and roots typical to FBR were observed, in a field trial of 36 onion cultivars during October 2020 in Bangalore, Karnataka. FBR incidence varied from 30-100% in this field (Fig 1 a-d). Symptomatic bulbs were washed with water, basal plate and fleshy leaves cut into 0.5 to 1 cm-size, surface disinfected with 1.5% sodium hypochlorite (NaOCl) for 3 min, and rinsed with sterile distilled water. Twenty pieces were placed on potato dextrose agar (PDA) in Petri plates and incubated at 25°C for 7 days. Colonies from single-spore isolates on PDA showed abundant white aerial mycelium. Colonies showed light pink or purple coloration on the reverse side of the culture plate with brown center (Fig 1e-f). Macroconidia were 19.13 to 28.35 (mean= 24.2) × 4.29 to 6.06 (mean= 5.05) µm, hyaline, falcate, with slightly curved apexes, and three to five septa. Microconidia were cylindrical to ellipsoid, aseptate, hyaline 8.20 to 12 (mean=10.0) × 3.55 to 4.79 (mean= 4.29) µm (Fig 1g). Chlamydospores were round, intercalary, hyaline, single or in chains (Fig 1h). Two isolates (IBFF-09 & IBFF-10) were analyzed for internal transcribed spacer-ITS (White et al. 1990) and translation elongation factor 1-α (tef1) gene (O'Donnell et al. 1998) by polymerase chain reaction (PCR) and sequenced. ITS and partial tef1 gene sequences of isolates IBFF-09 and IBFF-10 were submitted to the NCBI database (GenBank accession # ON394614&ON026859; # ON409480, ON093166 respectively). Phylogenetic analysis of tef1 gene placed the isolates with F. falciforme (Fig 1i). A pathogenicity test was performed by dipping roots of 28 days old healthy onion seedlings of a susceptible genotype 16/7Y GR3 into a conidial suspension (1 × 104 conidia/ml) of isolate IBFF-10 for 15 min and then transplanting the plants into pots containing sterilized potting mix. Inoculated plants developed typical symptoms of FBR and were all dead by 20 days post inoculation (Fig 1j) while the non-inoculated controls remained healthy. Pathogen was re-isolated from infected plants and showed the same morphology, ITS and tef1 sequence similarity as the original isolate, thus fulfilling Koch's postulates. Basal rot of onion by F. falciforme is reported from Mexico (Tirado-Ramírez et al. 2021). Till date, only F .oxysporum, F. proliferatum and F. solani have been implicated in onion FBR in India (Lee et al. 2021; Rathore and Patil, 2019). F. falciforme however, i prevalent in India and is reported to infect other crops (Gangaraj et al. 2022; Gupta et al. 2019; Homa et al. 2018). There is a high probability that this pathogen is contributing significantly to basal rot disease but it has not been reported yet. To our knowledge, this is the first report of F. falciforme infecting onions in India. In order to develop FBR resistant onion cultivars it is critical to identify and study the response of onion genotypes to different Fusarium spp causing the disease.

Dominance of resistance-breaking cotton leaf curl Burewala virus (CLCuBuV) in northwestern India.

Cotton leaf curl disease (CLCuD) is a major limitation to cotton production on the Indian subcontinent. A survey for viruses causing CLCuD was conducted during the 2009 and 2010 cropping seasons in the northwestern Indian cotton-growing belt in the states of Punjab, Haryana and Rajasthan. Partial sequences of 258 and full-length sequences of 22 virus genomes were determined. This study shows that the resistance-breaking cotton leaf curl Burewala virus (CLCuBuV) is now the dominant virus in many fields. The spread and establishment of the mutant CLCuBuV in northwestern India, the variation in its genomic sequence, its virulence and infectivity, and the implications for cotton breeding are discussed.

Virus-induced gene silencing.

In the postgenomic era, large-scale functional genomic approaches are necessary for converting sequence information into functional information. A para-genetic approach, called virus-induced gene silencing (VIGS), offers a rapid means of gaining insight into gene function in plants. VIGS system could be used to suppress endogenous gene expression by infecting plants with a recombinant virus vector (VIGS vector) carrying host-derived sequence. Here, we describe the use of tobacco rattle virus (TRV)-based VIGS technique to study gene function in Nicotiana benthamiana and tomato.

Induced ER chaperones regulate a receptor-like kinase to mediate antiviral innate immune response in plants.

Mounting an effective innate immune response against pathogens requires the rapid and global reprogramming of host cellular processes. Here we employed complementary proteomic methods to identify differentially regulated proteins early during a plant's defense response. Besides defense-related proteins, constituents of the largest category of upregulated proteins were cytoplasmic- and ER-residing molecular chaperones. Investigating the significance of upregulated ER chaperones, we find that silencing of ER-resident protein disulfide isomerases NbERp57 and NbP5 and the calreticulins NbCRT2 and NbCRT3 led to partial loss of N immune receptor-mediated defense against Tobacco mosaic virus (TMV). Furthermore, NbCRT2 and NbCRT3 were required for the expression of a previously uncharacterized induced receptor-like kinase (IRK). IRK is a plasma membrane-localized protein required for N-mediated hypersensitive response, programmed cell death, and resistance to TMV. These data support a model in which ER-resident chaperones are required for the accumulation of membrane-bound or secreted proteins during plant innate immunity.

Study of Arabidopsis thaliana resistome in response to cucumber mosaic virus infection using whole genome microarray.

The plant innate immune response is mediated by resistance (R) genes and involves hypersensitive response (HR) cell death. During resistance responses, the host undergoes net changes in the transcriptome. To understand these changes, we generated a whole genome transcript profile for RCY1-mediated resistance to cucumber mosaic virus strain Y (CMV-Y) in Arabidopsis. Using a very stringent selection criterion, we identified 444 putative factors belonging to nine different functional classes that show significant transcript regulation during Arabidopsis-CMV-Y interaction. Genes with unknown function formed the largest class. Other functional classes represented in the resistome include kinases and phosphatases, protein degradation machinery/proteases, transcriptional regulators, and others. Interestingly, several of the unknown function genes possess well characterized domains and secondly many genes encode small peptides with less than 100 amino acids. Analysis of 1.1 kb promoter regions of the 444 genes revealed that 9 out of the 12 known cis-binding elements are significantly associated with pathogen responsive cluster. Location and distribution of five prominent binding elements for select group of disease resistance related and unknown function genes is presented. The analysis also revealed 80 defense-responsive genes that might participate in R gene-mediated defense against both viral and bacterial pathogens. In addition, chromosome distribution of genes that respond to bacterial and viral pathogens suggests that they are located in small gene clusters and may be transcriptionally co-regulated. Exploring the precise function of the new genes identified in this analysis will offer new insights into plant defense.

The tobacco mosaic virus resistance gene, N.

Summary In this mini review we discuss recent advances in the understanding of the N gene-mediated resistance to tobacco mosaic virus (TMV). The tobacco N gene belongs to toll-interleukin-1 receptor homology/nucleotide binding/leucine rich repeat (TIR-NB-LRR) class of resistance genes. It encodes two transcripts, N(S) and N(L), by alternative splicing, both of which are required to confer resistance to TMV. The structure-function analysis of the N gene indicates that the TIR, NB and LRR domains are indispensable for its function. The N gene response is elicited by the C-terminal helicase domain of the 126 kDa TMV replicase protein. Tobacco N gene can also confer resistance to TMV in heterologous plants like tomato and Nicotiana benthamiana. Recent studies on N-mediated signalling suggest that EDS1, Rar1 and NPR1 genes play an important role in TMV resistance. Finally, we discuss current status of the N-mediated signal transduction and speculate directions for future work to understand N-TMV interaction.