A high-throughput protocol for testing heat-stress tolerance in pollen  

作　　者：Chenchen Zhao Abu Bakar Siddique Ce Guo Sergey Shabala Chengdao Li Zhonghua Chen Rajeev Varshney Meixue Zhou 

机构地区：[1]Tasmanian Institute of Agriculture,University of Tasmania,Prospect,TAS 7250,Australia [2]International Research Centre for Environmental Membrane Biology,Foshan University,Foshan 528000,China [3]School of Biological Science,University of Western Australia,Crawley 6009,Australia [4]Food Futures Institute,School of Agriculture,Western Crop Genetics Alliance,Murdoch University,Murdoch,WA 6150,Australia [5]Agriculture and Food,Department of Primary Industries and Regional Development,South Perth,WA 6150,Australia [6]School of Science,Western Sydney University,Penrith,NSW 2751,Australia [7]Hawkesbury Institute for the Environment,Western Sydney University,Penrith 2751,Au

出　　处：《aBIOTECH》2025年第1期63-71,共9页生物技术通报(英文版)

基　　金：supported by the Grains Research and Development Corporation(WSU2303-001RTX)of Australia.

摘　　要：Viable pollen is crucial for fertilization,but pollen is generally highly susceptible to heat stress.A quick,reliable method for testing the heat-stress tolerance of pollen is needed to improve the heat-stress tolerance in plants,but current methods require considerable space and labor.In addition,many such methods only test tolerance to a single constant temperature,making it time-consuming to screen heat tolerance over a wide temperature range and to examine the dynamics of pollen viability at different temperatures.To address this issue,we aimed to:(1)develop an easy,reliable method for measuring pollen viability at different temperatures;and(2)identify the best temperature range for screening pollen with high heat-stress tolerance.We harvested mature pollen from wheat(Triticum aestivum)plants and transferred it to a 96-well plate filled with liquid medium containing sucrose.We placed the plate in a PCR machine operating under a gradient PCR program to simultaneously test a range of temperatures.After incubating the pollen for 4 h,at temperatures ranging from 21.9 to 47℃,we examined the pollen grains under a light microscope and employed a specific image analysis pipeline to assess the effects of temperature on pollen morphology,germination,and tube growth.This method facilitated the high-throughput screening of many pollen samples,enabling rapid,reliable,and precise analysis of pollen viability in response to temperature.Our approach should be applicable to other plant species and could be used to identify quantitative trait loci or genes influencing heat stress tolerance in pollen for breeding programs.

关 键 词：Heat stress Pollen germination Pollen tube development Triticum aestivum L. Hordeum vulgare L. 

分 类 号：Q945.4[生物学—植物学]