Simple cryopreservation protocol for Luffa pollen: enhancing breeding efficiency

Arvind Nagar; Ravi Gowthami; Amish Kumar Sureja; Anilabha Das Munshi; Manjusha Verma; Awani Kumar Singh; Niharika Mallick; Jogendra Singh; Subhash Chander; Muthusamy Shankar; Pooja Pathania; Subramani Rajkumar

doi:10.3389/fpls.2023.1268726

Simple cryopreservation protocol for Luffa pollen: enhancing breeding efficiency

Front Plant Sci. 2023 Oct 27:14:1268726. doi: 10.3389/fpls.2023.1268726. eCollection 2023.

Authors

Affiliations

¹ Krishi Vigyan Kendra (KVK), Jhalawar, Agricultural University, Kota, India.
² Division of Germplasm Conservation, ICAR-National Bureau of Plant Genetic Resources, New Delhi, India.
³ Division of Vegetable Science, ICAR-Indian Agricultural Research Institute, New Delhi, India.
⁴ Division of Genomic Resources, ICAR-National Bureau of Plant Genetic Resources, New Delhi, India.
⁵ CPCT, ICAR-Indian Agricultural Research Institute, New Delhi, India.
⁶ Division of Genetics, ICAR-Indian Agricultural Research Institute, New Delhi, India.
⁷ Division of Plant Genetic Resources, The Graduate School, Indian Agricultural Research Institute, Indian Council of Agricultural Research (ICAR), New Delhi, India.

^# Contributed equally.

Abstract

This study aimed to develop a long-term pollen storage protocol for Luffa species (L. acutangula, L. cylindrica, L. echinata, and L. graveolens) and assess its potential for crop improvement. The optimal medium for in vitro pollen germination varied by species, with Brewbaker and Kwack (BK) medium with 10% sucrose suitable for L. acutangula, L. cylindrica, and L. echinata, and BK medium with 3% sucrose ideal for L. graveolens. Overestimation in staining tests compared to in vitro pollen germination was observed. The best results for cryopreservation were achieved with desiccation periods of 20, 30, and 40 min, maintaining moisture content between 14.04% and 18.55%. Pollen viability was negatively correlated with storage temperature (25, 4, and -20°C) and duration. Cryopreserved pollen at -196°C exhibited the highest viability over a prolonged period (2 months) and was comparable to fresh pollen in terms of germination, ovule fertilization, and fruit and seed set. This study presents a simple and reproducible pollen cryopreservation protocol applicable across Luffa species, facilitating long-term storage and its use in crop improvement efforts.

Keywords: Luffa; cryopreservation; incubation temperature; long-term storage; pollen; storage temperature; ultra-low freezing.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. The authors are thankful to the ICAR-Indian Agricultural Research Institute, New Delhi for providing financial support and conduct of the research program of the Ph.D student, Mr. AN. The authors are thankful to the World Bank–ICAR-funded NAHEP-CAAST program of CSK-HPKV Palampur and CPCT, IARI for funding the research program.