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Validation of molecular response of tuberization in response to elevated temperature by using a transient Virus Induced Gene Silencing (VIGS) in potato

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Abstract

Temperature plays an important role in potato tuberization. The ideal night temperature for tuber formation is ~17 °C while temperature beyond 22 °C drastically reduces the tuber yield. Moreover, high temperature has several undesirable effects on the plant and tubers. Investigation of the genes involved in tuberization under heat stress can be helpful in the generation of heat-tolerant potato varieties. Five genes, including StSSH2 (succinic semialdehyde reductase isoform 2), StWTF (WRKY transcription factor), StUGT (UDP-glucosyltransferase), StBHP (Bel1 homeotic protein), and StFLTP (FLOWERING LOCUS T protein), involved in tuberization and heat stress in potato were investigated. The results of our microarray analysis suggested that these genes regulate and function as transcriptional factors, hormonal signaling, cellular homeostasis, and mobile tuberization signals under elevated temperature in contrasting KS (Kufri Surya) and KCM (Kufri Chandramukhi) potato cultivars. However, no detailed report is available which establishes functions of these genes in tuberization under heat stress. Thus, the present study was designed to validate the functions of these genes in tuber signaling and heat tolerance using virus-induced gene silencing (VIGS). Results indicated that VIGS transformed plants had a consequential reduction in StSSH2, StWTF, StUGT, StBHP, and StFLTP transcripts compared to the control plants. Phenotypic observations suggest an increase in plant senescence, reductions to both number and size of tubers, and a decrease in plant dry matter compared to the control plants. We also establish the potency of VIGS as a high-throughput technique for functional validation of genes.

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Acknowledgements

We are thankful to the Indian Council of Agricultural Research, New Delhi, for their financial support and ICAR-Central Potato Research Institute (CPRI), Shimla, Himachal Pradesh, for their help in execution and monitoring the experiment.

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  1. Maharishi Tomar, Sundaresha, S. and Baljeet Singh contributed equally to this work.

Authors and Affiliations

  1. ICAR-Central Potato Research Institute, Shimla, H.P., 171001, India

    Maharishi Tomar, Sundaresha S., Baljeet Singh, Vinay Bhardwaj, Salej Sood, Brajesh Singh, Neha Salaria, Kajal Thakur, Ashwani Kumar & Neha Sharma

  2. ICAR-Indian Grassland and Fodder Research Institute, Jhansi, U.P., 284003, India

    Maharishi Tomar

  3. Lovely Professional University, Jalandhar, Punjab, 144411, India

    Neha Salaria, Kajal Thakur & Umesh Goutam

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  1. Maharishi Tomar
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Contributions

MT developed the VIGS constructs. SS, KT, and NS raised the plant material. SS, BLS, KT, NS, and AK contributed to physiological studies and molecular characterization and carried out all the wet lab experiments. SS, VB, SSD, and UG analyzed the data. SS, VB, SSD, and BS mentored the whole study. SS, MT, and BLS wrote the MS. All authors have read and approved the MS.

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Correspondence to Sundaresha S..

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Supplementary Information

ESM 1

Supporting Figure. S1 Graphical illustration of pTZ57R/T (A) and pTRV2 (B) with genes of interest at MCS. (PNG 122687 kb)

High Resolution (TIF 1963 kb)

ESM 2

(PNG 125634 kb)

High Resolution (TIF 2073 kb)

ESM 3

Supporting Sequence information 1-5: Graphical representation of CDS region of selective gene segment showing the siRNA candidate fragments (DOCX 632 kb)

Supporting Table S1

Details of genes used in the study and their PGSC ID (DOCX 13 kb)

Supporting Table S2

Position of selected CDS regions for VIGS study, based on siRNA target finder (DOCX 14 kb)

Supporting Table S3

Plant Dry weight ratio in VIGS treated and control KS plants. (DOCX 13 kb)

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Tomar, M., S., S., Singh, B. et al. Validation of molecular response of tuberization in response to elevated temperature by using a transient Virus Induced Gene Silencing (VIGS) in potato. Funct Integr Genomics 21, 215–229 (2021). https://doi.org/10.1007/s10142-021-00771-2

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