Development of in vitro germination and cryopreservation protocols of pollen in wild Lathyrus species to overcome longevity and asynchronous flowering constraints

Rinky Panda, Padmavati G. Gore, Aishmita Gantait, Esha Singh, Mahesh Rao, S. Rajkumar, Shiv Kumar Agrawal, Kuldeep Tripathi, Sandhya Gupta. (10/8/2026). Development of in vitro germination and cryopreservation protocols of pollen in wild Lathyrus species to overcome longevity and asynchronous flowering constraints. Plant Cell, Tissue and Organ Culture, 166.

Abstract

The genus Lathyrus comprises several important multipurpose and resilient legume species, serving as a valuable reservoirof genetic diversity for stress tolerance and nutritional enhancement. However, its wild species are underutilized in breeding due to asynchronous flowering, short pollen longevity, and lack of standardized germination and conservation protocols. The present study is a novel attempt to optimize the in vitro pollen germination protocol and to develop a reliable cryopreservation strategy for six Lathyrus species (L. pseudocicera, L. odoratus, L. tingitanus, L. annuus, L. aphaca and L. cicera). Pollen exhibited strong species-specific responses to sucrose concentration, with maximum germination recorded on 15% sucrose in L. pseudocicera (84.7%) and L. tingitanus (78.3%), on 20% sucrose in L. annuus (81.9%) and L. aphaca (76.1%), on 25% sucrose in L. odoratus (86.8%), and on 30% sucrose in L. cicera (65.1%). Fresh pollen showed high viability (65.1–86.8%) but declined sharply after LN exposure, losing viability by 24 h. Desiccated pollen of L. pseudocicera showed the highest LN survival (82.0%) after 20 min of desiccation, while L. odoratus and L. tingitanus peaked at 25–30 min (77.6%), L. annuus and L. aphaca at 5 min (71.6–73.6%), and L. cicera at 15 min (57.3%). Cryopreserved pollen successfully produced normal pods and seed setting with normal seed germination at par with the control seed lot. This is the first systematic report on in vitro pollen germination and cryopreservation in wild Lathyrus species, providing a practical framework to overcome asynchronous flowering, improve pollen longevity, and enhance germplasm conservation and interspecific hybridization in breeding programs.