Establishment of effective plantlets regeneration protocol via isolated microspore culture in Malaysian indica rice MR219

Rahman, Zuraida Abdul; Seman, Zulkifli Ahmad; Othman, Ayu Nazreena; Ghaffar, Mohamad Bahagia Ab; Razak, Shahril Ab; Yusof, Muhammad Fairuz Mohd; Nasir, Khairun Hisam; Ahmad, Khalil; Chow, Yeow Lit; Teo, Chee How; Saad, Norsharina Md; Subramaniam, Sreeramanan

doi:10.1007/s11816-022-00742-4

Cited by 3 publications

(1 citation statement)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thousands of embryogenic calli can thus be rapidly induced, many of which can later develop into regenerated plants (Li and Devaux 2005 ; Esteves et al 2014 ). In recent years, higher frequencies of embryogenic callus formation and green plantlet regeneration have been achieved in several plant species, such as rice ( Oryza sativa ), wheat ( Triticum aestivum ), tobacco ( Nicotiana tabacum ), rapeseed, and eggplant ( Solanum melongena ) (Shariatpanahi et al 2006 ; Mohammadi et al 2012 ; Shariatpanahi and Ahmadi 2016 ; Dong et al 2021 ; Calabuig-Serna et al 2022 ; Rahman et al 2022 ). An efficient isolated microspore culture system was established in barley ( Hordeum vulgare ) following the optimization of factors that affect microspore embryogenesis (Lu et al 2008 , 2016 ).…”

Section: Introductionmentioning

confidence: 99%

Generating homozygous mutant populations of barley microspores by ethyl methanesulfonate treatment

et al. 2023

View full text Add to dashboard Cite

Induced mutations are important for genetic research and breeding. Mutations induced by physical or chemical mutagenesis are usually heterozygous during the early generations. However, mutations must be fixed prior to phenotyping or field trials, which requires additional rounds of self-pollination. Microspore culture is an effective method to produce double-haploid (DH) plants that are fixed homozygotes. In this study, we conducted ethyl methanesulfonate (EMS)-induced mutagenesis of microspore cultures of barley (Hordeum vulgare) cultivar ‘Hua30’ and landrace ‘HTX’. The EMS concentrations were negatively correlated with the efficiency of callus induction and the frequency of mutant plant regeneration. The two genotypes showed different regeneration efficiencies. The phenotypic variation of the regenerated M1 plants and the presence of genome-wide nucleotide mutations, revealed by whole-genome sequencing, highlight the utility of EMS-induced mutagenesis of isolated microspore cultures for developing DH mutants. Genome-wide analysis of the mutation frequency in the regenerated plants revealed that a considerable proportion of mutations resulted from microspore culture (somaclonal variation) rather than EMS-induced mutagenesis. In addition to producing a population of 1972 homozygous mutant lines that are available for future field trials, this study lays the foundation for optimizing the regeneration efficiency of DH plants and the richness of mutations (mainly by fine-tuning the mutagen dosage).

show abstract

Section: Introductionmentioning

confidence: 99%

Generating homozygous mutant populations of barley microspores by ethyl methanesulfonate treatment

et al. 2023

View full text Add to dashboard Cite

show abstract

Transcriptomics integrated with metabolomics reveals the effect of cold stress on rice microspores

Li,

Zong,

et al. 2023

BMC Plant Biol

View full text Add to dashboard Cite

Background Microspore culture is one of the important biotechnological tools in plant breeding. The induction of microspore embryogenesis is a critical factor that affects the yield of microspore-derived embryo productions. Cold treatment has been reported to reprogram the gametophytic pathway in various plant species. However, the exact mechanism(s) underlying the effect of cold pre-treatment of floral buds on the efficiency of ME is still not clear. Results In this study, the effects of cold stress on the microspore totipotency of rice cultivar Zhonghua 11 were investigated. Our results revealed that a 10-day cold treatment is necessary for microspore embryogenesis initiation. During this period, the survival rate of microspores increased and reached a peak at 7 days post treatment (dpt), before decreasing at 10 dpt. RNA-seq analysis showed that the number of DEGs increased from 3 dpt to 10 dpt, with more downregulated DEGs than upregulated ones at the same time point. GO enrichment analysis showed a shift from ‘Response to abiotic stimulus’ at 3 dpt to ‘Metabolic process’ at 7 and 10 dpt, with the most significant category in the cellular component being ‘cell wall’. KEGG analysis of the pathways revealed changes during cold treatment. Mass spectrometry was used to evaluate the variations in metabolites at 10 dpt compared to 0 dpt, with more downregulated DEMs being determined in both GC-MS and LC-MS modes. These DEMs were classified into 11 categories, Most of the DEMs belonged to ‘lipids and lipid-like molecules’. KEGG analysis of DEMs indicates pathways related to amino acid and nucleotide metabolism being upregulated and those related to carbohydrate metabolism being downregulated. An integration analysis of transcriptomics and metabolomics showed that most pathways belonged to ‘Amino acid metabolism’ and ‘Carbohydrate metabolism’. Four DEMs were identified in the interaction network, with stearidonic acid involving in the most correlations, suggesting the potential role in microspore totipotency. Conclusions Our findings exhibited the molecular events occurring during stress-induced rice microspore. Pathways related to ‘Amino acid metabolism’ and ‘Carbohydrate metabolism’ may play important roles in rice microspore totipotency. Stearidonic acid was identified, which may participate in the initiation of microspore embryogenesis.

show abstract

Efficient isolated microspore culture protocol for callus induction and plantlet regeneration in japonica rice (Oryza sativa L.)

Gao,

Zong,

Zhang

et al. 2024

Plant Methods

View full text Add to dashboard Cite

Background Isolated microspore culture is a useful biotechnological technique applied in modern plant breeding programs as it can produce doubled haploid (DH) plants and accelerate the development of new varieties. Furthermore, as a single-cell culture technique, the isolated microspore culture provides an excellent platform for studying microspore embryogenesis. However, the reports on isolated microspore culture are rather limited in rice due to the low callus induction rate, poor regeneration capability, and high genotypic dependency. The present study developed an effective isolated microspore culture protocol for high-frequency androgenesis in four japonica rice genotypes. Several factors affecting the isolated microspore culture were studied to evaluate their effects on callus induction and plantlet regeneration. Results Low-temperature pre-treatment at 4 ℃ for 10–15 days could effectively promote microspore embryogenesis in japonica rice. A simple and efficient method was proposed for identifying the microspore developmental stage. The anthers in yellow-green florets located on the second type of primary branch on the rice panicle were found to be the optimal stage for isolated microspore culture. The most effective induction media for callus induction were IM2 and IM3, depending on the genotype. The optimal concentration of 2, 4-D in the medium for callus induction was 1 mg/L. Callus induction was negatively affected by a high concentration of KT over 1.5 mg/L. The differentiation medium suitable for japonica rice microspore callus comprised 1/2 MS, 2 mg/L 6-BA, 0.5 mg/L NAA, 30 g/L sucrose, and 6 g/L agar. The regeneration frequency of the four genotypes ranged from 61–211 green plantlets per 100 mg calli, with Chongxiangjing showing the highest regeneration frequency. Conclusions This study presented an efficient protocol for improved callus induction and green plantlet regeneration in japonica rice via isolated microspore culture, which could provide valuable support for rice breeding and genetic research.

show abstract

Establishment of effective plantlets regeneration protocol via isolated microspore culture in Malaysian indica rice MR219

Cited by 3 publications

References 27 publications

Generating homozygous mutant populations of barley microspores by ethyl methanesulfonate treatment

Generating homozygous mutant populations of barley microspores by ethyl methanesulfonate treatment

Transcriptomics integrated with metabolomics reveals the effect of cold stress on rice microspores

Efficient isolated microspore culture protocol for callus induction and plantlet regeneration in japonica rice (Oryza sativa L.)

Contact Info

Product

Resources

About