Actinidia chinensis is a commercially important fruit, and tetraploid breeding of A. chinensis is of great significance for economic benefit. In order to obtain elite tetraploid cultivars, tetraploid plants were induced by colchicine treatment with leaves of diploid A. chinensis ‘SWFU03’. The results showed that the best treatment was dipping leaves 30 h in 60 mg l−1 colchicine solutions, with induction rate reaching 26%. Four methods, including external morphology comparison, stomatal guard cell observation, chromosome number observation and flow cytometry analysis were used to identify the tetraploid of A. chinensis. Using the induction system and flow cytometry analysis methods, 187 tetraploid plants were identified. Three randomly selected tetraploid plants and their starting diploid plants were further subjected to transcriptome analysis, real-time quantitative polymerase chain reaction (RT-qPCR) and methylation-sensitive amplification polymorphism (MSAP) analysis. The transcriptome analysis results showed that there were a total of 2230 differentially expressed genes (DEG) between the diploid and tetraploid plants, of which 660 were downregulated and 1570 upregulated. The DEGs were mainly the genes involved in growth and development, stress resistance and antibacterial ability in plants. RT-qPCR results showed that the gene expression levels of the growth and stress resistance of tetraploid plants were higher than those of diploid ones at the transcriptome level. MSAP analysis of DNA methylation results showed that tetraploid plants had lower methylation ratio than diploid ones. The present results were valuable to further explore the epigenetics of diploid and tetraploid kiwifruit plants.
Yellow kiwifruit ( Actinidia chinensis ) is highly susceptible to severe weather, such as low temperature and frost, which may affect the production in the coming year. And the cold-resistant mechanism of kiwifruit associated with gene regulation is poorly investigated. To botain cold-resistant germplam, to provide insight into the causes of differences in low temperature tolerance due to ploidy and to better understand cold-adaptive mechanisms in tetraploid kiwifruit, the diploid yellow kiwifruit ‘SWFU03’ and its tetraploid plantlets were subjected to cold-tolerant screening with L-hydroxyproline (L-Hyp) and low temperature, the selected ones were then analyzed by transcriptome data and confirmed by RT-qPCR. The results showed that the survival rate of tetraploid plants was 62.22% when treated with 8 mmol/L L-Hyp for 30 days, while all the diploid ones died. After treated with 0°C for 12 h, then at room temperature for seven days, the survival rate of tetraploid plantlets was 42.22%, while all diploidy died. Hence, cold tolerance of the tetraploid plantlets was stronger than that of the diploid genotypes. Using these two screening systems, 126 cold-resistant tetraploid tissue culture plantlets were obtained. A total of 1630 differentially expressed genes (DEGs) were identified, of which 619 were up-regulated and 1011 were down-regulated in the low temperature treatment goup. The DEGs enriched in the cold-tolerance related pathways mainly included plant hormone signal transduction, and starch and sucrose metabolism pathway. RT-qPCR analysis confirmed the expression levels of eight up-regulated genes in these pathways in the cold-resistant mutants. In conclusion, this study has identified cold-resistant yellow kiwifruit plantlets and cold-tolerance related genes. Moreover, the dataset got in this study advances our knowledge of the cold-adaptive genes in the regulatory networks and leads to understand the cold tolerance mechanisms in the tetraploid yellow kiwifruit.
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