Abstract
Drought causes severe damage to sugarcane, reducing its product yield. Given Thailand’s weather conditions and topography, a breeding program to develop new sugarcane genotypes with a high tolerance for water stress is important to the sugarcane industry. This study created new water stress tolerant sugarcane genotypes using ethyl methanesulfonate (EMS) mutagenesis in the sugarcane cultivar Khon Kaen 3. Using 16 mM of EMS for 4 h induced callus mutagenesis (survival rate, 57.5%). The survival rates of calli treated with 10 mM of EMS for 2 and 4 h in selective media with 15% PEG were higher than that of non-EMS-treated calli. The selected calli survived and grew on selective media with 20% PEG, while non-EMS-treated calli did not grow. The mutant plantlets developed from EMS-treated calli on selective media with 20% PEG for 4 weeks had varying survival rates: 72.25% (10 mM of EMS for 2 h), 75.85% (10 mM of EMS for 4 h), and 60.61% (16 mM of EMS for 4 h). Both healthy mutant sugarcane plants (2,086) and non-mutant plants (234) were cultured on the media with 20% PEG for 16 weeks. Of these, 462 mutant sugarcane plantlets survived and developed on the media, but all the non-mutant sugarcane plantlets died during the selection process. Mutagenesis induced using treatment 4 produced the highest frequency of mutant sugarcane plantlets with water-stress tolerance (45.5%). In total, 136 selected mutant sugarcane plants were transplanted to a greenhouse for evaluation under managed water stress. Fourteen mutant sugarcane plants stayed green after the third cycle of water stress, but the KK3 sugarcane cultivar showed damage on 50% of the leaves. Thus, EMS mutagenesis and evaluation using in vitro and greenhouse methods were successful in developing new sugarcane clones with high water-stress tolerance, which is important for sugarcane breeding programs.
Highlights
Sugarcane is an economic crop that has a substantial impact on the economy of ailand, where it is the primary raw material for the production of sugar used in many highvalue products and downstream industries, including bioethanol and electricity production [1]
Sterile shoot tips of Khon Kaen 3 (KK3) were cut into slices approximately 3–5 mm thick and placed on Murashige and Skoog (MS) medium supplemented with three different treatments: 3 mg/l 2,4-D, 3 mg/l 2,4D + 0.5 mg/l benzylamino purine (BAP), or 3 mg/l 2,4-D + 10% coconut water. e sterilized explants were incubated in the dark at 25°C ± 2°C
Callus formation was discovered on leaf tissue after 7 days of culture on the callus induction media. e MS media supplemented with 3 mg/l of 2,4-D produced the highest rate of callus initiation (96.43%); the second-highest rate was observed in MS media supplemented with 3 mg/l of 2,4-D plus 15% coconut water (74.07%), as shown in Table 1. e lowest rate of callus induction was produced using the MS media supplemented with 3 mg/l of 2,4-D, and 0.5 mg/l of BAP (9.47%)
Summary
Sugarcane is an economic crop that has a substantial impact on the economy of ailand, where it is the primary raw material for the production of sugar used in many highvalue products and downstream industries, including bioethanol and electricity production [1]. Drought is adversely affecting many of ailand’s economic crops, including sugarcane. Cane production in 2020 was dropped approximately 43%, compared with the previous year, because of the drought [2]. Drought often occurs in the middle of ailand’s sugarcane planting season, often having a devastating effect on yield as it reduces the number of shoots per clump and stunts the sugarcane stalks. Sugarcane farmers cannot avoid this annual period of drought because sugarcane grows over a span of 12–15 months. Sugarcane farmers cannot avoid this annual period of drought because sugarcane grows over a span of 12–15 months. erefore, a systematic breeding program
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