Reddy, M. Gurivi

A field investigation was conducted during the winter seasons of 2021–22 and 2022–23 at the Jawaharlal Nehru Krishi Vishwa Vidyalaya (JNKVV), Jabalpur, Central India, to identify and characterize the phytoplasma strains associated with niger (var. JNC6) and sesame (var. RT315) exhibiting severe phyllody symptoms. Disease incidence ranged from 0-3%, and 3 to 15% were recorded in niger and sesame crops during the study period. Nested PCR assays using universal phytoplasma primer pairs P1/P7 followed by R16F2n/R16R2 consistently amplified ~1.25 kb fragments of the 16S rRNA gene from both the symptomatic plants, confirming phytoplasma association. Additional amplification of phytoplasma-specific sec A (~600 bp) and sec Y (~1.7 kb) gene fragments further validated the phytoplasma infection in both crops. Two predominant leafhopper species, Amrasca biguttula and Orosius albicinctus , colonizing and feeding niger and sesame fields were identified. Notably, only Amrasca biguttula was tested positive for phytoplasma using the similar PCR assays mentioned above, implicating it as a putative vector. Sequence identity, phylogenetic analysis, and in silico RFLP profiling of 16S rRNA, sec A, and sec Y gene sequences from both the symptomatic plants and leafhoppers confirmed the presence of a ‘ Ca. P. asiaticum’ related strain (16SrII-C subgroup). This study represents the first global report of 16SrII-C subgroup phytoplasma infection in niger, highlighting a new host record. The detection of an identical phytoplasma strain in niger, sesame, and Amrasca biguttula suggests a possible epidemiological link and underscores the role of these natural hosts and the leafhopper in the dissemination of ‘ Ca. P. asiaticum’-related phytoplasma strains in Central India.

Background: Alfalfa, Medicago sativa L. is the most important and widely grown leguminous fodder crop in temperate and tropical regions of the world. The production of alfalfa crop is limited by several biotic stresses, among which witches’ broom disease (AWB) was reported to cause significant economic losses. Methods: The phytoplasma infected alfalfa plants were collected from a central research farm, ICAR-IGFRI, Jhansi, U.P. Qualitative parameters such as crude protein content, acid detergent fibre and neutral detergent fibre were estimated in diseased and healthy plants. Phytoplasma universal primer (P1/P7) and nested primer (R16mF2/R16mR1) were used for the molecular characterization of AWB infected plants and phytoplasma infected, Parthenium hysterophorus. Result: The incidence of AWB disease ranged from 8-10%. The quantitative analysis of disease plants showed reduced plant height (-35%), fresh weight (-46.89%) and dry weight (-50.08%) compared to healthy plants. The diseased plant recorded low crude protein content (-21.38%) and higher dry matter content (+0.68%), acid detergent fibre (+33.72%) and neutral detergent fibre (+13.06%). The association of phytoplasma in diseased alfalfa and parthenium samples was confirmed by using P1/P7 and R16mF2/R16mR1 primer pair and Blastn analysis shared 99.6-100% similarity with ‘Candidatus Phytoplasma australasia’ belongs to the 16Sr group II-D.