Abstract
China has abundant of local citrus resources, which had many seeds. The Seedless is the aim of the local citrus breeding. Triploid citurs resources usually obtained by crosssing breeding between the diploid and tetraploid plants. However, the resources of tetraploid citrus are limited. It has been reported that the hybrid between diploid and diploid plants can produce triploid citrus, which not only saves the breeding time, but also shortens the breeding cycles. In this project, flow cytometry detection assisted by embryo rescue system and SSR identification technology were used to improve the probability of obtaining polyploidy from the hybrid offspring, and provide technical support for obtaining polyploidy from large-scale hybridization between diploid and diploid. plants. Three diploid pollen cultivars were crossbred with Hunan local cultivar 8306, immature seeds were collected for embryo rescue to obtain regenerative plants, plant ploidy was detected by flow cytometry and stoma electron microscope, and offspring genotypes were identified by SSR. After 100 d hybrid between three diploid pollens and Ponkan 8 306, 405 plants were obtained by embryo rescue technique. And 70 plants survived in the greenhouse after transplantation. Through flow cytometry instrument and stomatal electron microscope inspection, the results showed that SSR analysis validated that there are 13 polyploid plants, including 1 tetraploid plant and 12 triploid plants. The ratio of polyploidy was 18.57%. This study is obtained triploid and tetraploid plants by Ponkan embryo rescue from diploid interspecific hybridization. In this study, the embryo rescue technology system and the early identification system for polyploid hybridization were established, and a lot of new polyploid germplasm were obtained.
Highlights
Triploid citurs resources usually obtained by crosssing breeding between the diploid and tetraploid plants
Three diploid pollen cultivars were crossbred with Hunan local cultivar 8306, immature seeds were collected for embryo rescue to obtain regenerative plants, plant ploidy was detected by flow cytometry and stoma electron microscope, and offspring genotypes were identified by SSR
The results showed that the probability of polyploidy was 18.57% by flow cytometry and leaf stomatal scanning electron microscopy
Summary
Triploid citurs resources usually obtained by crosssing breeding between the diploid and tetraploid plants. It has been reported that the hybrid between diploid and diploid plants can produce triploid citrus, which saves the breeding time, and shortens the breeding cycles In this project, flow cytometry detection assisted by embryo rescue system and SSR identification technology were used to improve the probability of obtaining polyploidy from the hybrid offspring, and provide technical support for obtaining polyploidy from large-scale hybridization between diploid and diploid. This study is obtained triploid and tetraploid plants by Ponkan embryo rescue from diploid interspecific hybridization. With ponkan as the female parent, we hope to use diploid and diploid hybrid breeding technology, assisted embryo rescue technology to improve the survival rate of immature embryos or abortive embryos, and use flow cytometry and stomatal scanning electron microscopy to identify the early ploidy of seedlings, so as to Molecular Plant Breeding 2020, Vol., No.28, 1-8 http://genbreedpublisher.com/index.php/mpb quickly obtain polyploid progenies, and use assisted SSR molecular markers to identify the homozygous genotype and heterozygous genotype. The embryonic rescue technique system and the early identification system of polyploid obtained by interspecific hybridization of polyploid citrus were established in this study, which could provide theoretical basis for the breeding of new seedless varieties and the basic research of hybridization
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