Pollen: A Potential Explant for Genetic Transformation in Wheat (Triticum aestivum L.)

Abstract

The use of biotechnology for the genetic improvement of wheat (Triticum aestivum L.) has been hampered by its recalcitrance to standard transformation and regeneration protocols. Male gametes present a potentially useful option for introducing gene edits via clustered regularly interspaced short palindromic repeats (CRISPR). However, the utility of male gametes for introducing genetic improvements would be dependent on the retention of viability after treatment to introduce the CRISPR components. We have studied wheat pollen morphology and its viability in a range of germination media to identify conditions that optimize the viability of in vitro hydrated pollen. The size, shape, and aperture from seven different wheat genotypes were compared using scanning electron microscope (SEM). The SEM results revealed that the pollen of all of the wheat genotypes examined in this study were monoporate; however, a significant variation in the size of the mature pollen grains was observed. The hydrated pollen of the wheat genotypes remained viable for up to five hours at 20 ± 2 °C. Of all of the germination media tested, the medium containing 5% sucrose, 10% PEG4000, 100 mg/L boric acid, 200 mg/L calcium nitrate, 100 mg/L potassium nitrate, and 100 mg/L magnesium sulphate at pH 6.5 achieved the highest percentage of pollen germination after 5 h of hydration. Impedance Flow Cytometry (IFC) provided similar results to the in vitro germination study. This work elucidates important factors that can form the basis for a pollen-based non-genetically modified system for gene editing in wheat.

Screening and optimisation of in vitro pollen germination medium for sweetpotato (Ipomoea batatas)

“…To screen a suitable initial medium formulation for in vitro sweetpotato pollen germination, extensive pre-experiments were conducted using many pollen germination media used in other plants. We screened 10 media used in different plants [ 40 – 49 ] (Table 1 , M1–M10) and simplified the composition of M1 (Table 1 , M11), which had the best germination efficiency in the pre-experiments. Pollen germination rates were compared between different media and pure water (Table 1 , CK), and the medium with the highest germination rate was selected for optimisation as the initial medium for in vitro sweetpotato pollen germination.…”

A systematic review of the role of integrated farming and the participation of universities in ensuring food security: Malaysia’s effort

“…One of the efforts made by universities to ensure food security is through research and development (Ezanee, 2022 ). Agricultural universities research new crop varieties (Glenn et al , 2017 ; Kanwal et al ., 2022 ; Gros-Balthazard et al ., 2023 ), innovative farming techniques (Depenbusch et al ., 2021 ; Liu et al ., 2021 ; Montes de Oca Munguia et al ., 2021 ), and sustainable agricultural practices to improve productivity (Piñeiro et al ., 2020 ; Muhie, 2022 ; Boufous et al ., 2023 ) and efficiency in food production (Bin Rahman and Zhang, 2022; Jiang et al , 2022 ). They also research climate change impacts on food production and develop strategies to adapt to changing conditions (Gedik and Günel, 2021 ; Filho et al ., 2022 ).…”

Дослідження Характеру Акумуляції Розподілу Вільного Проліну В Органах Рослин За Умов Норми Та Стресу

“…Для отримання нових стрес-стійких форм рослин активно застосовують біотехнологічні методи, у тому числі генетичну інженерію, однією із складових якої є Agrobacterium-опосередкована трансформація in vitro та in planta. Це новітній метод, що передбачає активне втручання в геном рослини через пряме перенесення природних (виділених із біологічних об'єктів) конкретних генів або штучно створених дизайнерських конструкцій [7,23,29,34].…”

“…Відомо, що в загальному випадку рівень вільного pro саморегулюється системою синтезу/деградації/транспорту [4,5,6,7,8,10,17]. Геном синтезу є Δ 1 -піролін-5-карбоксилатсинтетаза, (П5КС); ген деградації проліндегідрогеназа, (ПДГ); транспортується pro системою власних та загальних транспортерів [7,9,13,23,35].…”

“…Фермент каталізує перетворення α-кетоглутарат → П5К. У літературі є відомості, що ген ОАТ може мати відношення до протистояння абіотичним стресам [5,7,17,23,34,35].…”