Paternal chromosome elimination of inducer triggers induction of double haploids in Brassica napus
A synthetic octoploid rapeseed, Y3380, induces maternal doubled haploids when used as a pollen donor to pollinate plant. However, the mechanism underlying doubled haploid formation remains elusive. We speculated that double haploid induction occurs as the inducer line’s chromosomes pass to the mater...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-10-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2023.1256338/full |
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| author | Shihui Zhao Shihui Zhao Liangjun Huang Liangjun Huang Qing Zhang Ying Zhou Ying Zhou Meicui Yang Meicui Yang Haoran Shi Yun Li Jin Yang Chao Li Xianhong Ge Wanzhuo Gong Jisheng Wang Qiong Zou Lanrong Tao Zeming Kang Zhuang Li Chaowen Xiao Qiong Hu Shaohong Fu Shaohong Fu |
| author_facet | Shihui Zhao Shihui Zhao Liangjun Huang Liangjun Huang Qing Zhang Ying Zhou Ying Zhou Meicui Yang Meicui Yang Haoran Shi Yun Li Jin Yang Chao Li Xianhong Ge Wanzhuo Gong Jisheng Wang Qiong Zou Lanrong Tao Zeming Kang Zhuang Li Chaowen Xiao Qiong Hu Shaohong Fu Shaohong Fu |
| author_sort | Shihui Zhao |
| collection | DOAJ |
| description | A synthetic octoploid rapeseed, Y3380, induces maternal doubled haploids when used as a pollen donor to pollinate plant. However, the mechanism underlying doubled haploid formation remains elusive. We speculated that double haploid induction occurs as the inducer line’s chromosomes pass to the maternal egg cell, and the zygote is formed through fertilization. In the process of zygotic mitosis, the paternal chromosome is specifically eliminated. Part of the paternal gene might have infiltrated the maternal genome through homologous exchange during the elimination process. Then, the zygote haploid genome doubles (early haploid doubling, EH phenomenon), and the doubled zygote continues to develop into a complete embryo, finally forming doubled haploid offspring. To test our hypothesis, in the current study, the octoploid Y3380 line was back bred with the 4122-cp4-EPSPS exogenous gene used as a marker into hexaploid Y3380-cp4-EPSPS as paternal material to pollinate three different maternal materials. The fertilization process of crossing between the inducer line and the maternal parent was observed 48 h after pollination, and the fertilization rate reached 97.92% and 98.72%. After 12 d of pollination, the presence of cp4-EPSPS in the embryo was detected by in situ PCR, and at 13–23 d after pollination, the probability of F1 embryos containing cp4-EPSPS gene was up to 97.27%, but then declined gradually to 0% at 23–33 d. At the same time, the expression of cp4-EPSPS was observed by immunofluorescence in the 3rd to 29th day embryo. As the embryos developed, cp4-EPSPS marker genes were constantly lost, accompanied by embryonic death. After 30 d, the presence of cp4-EPSPS was not detected in surviving embryos. Meanwhile, SNP detection of induced offspring confirmed the existence of double haploids, further indicating that the induction process was caused by the loss of specificity of the paternal chromosome. The tetraploid-induced offspring showed infiltration of the induced line gene loci, with heterozygosity and homozygosity. Results indicated that the induced line chromosomes were eliminated during embryonic development, and the maternal haploid chromosomes were synchronously doubled in the embryo. These findings support our hypothesis and lay a theoretical foundation for further localization or cloning of functional genes involved in double haploid induction in rapeseed. |
| first_indexed | 2024-03-11T14:52:50Z |
| format | Article |
| id | doaj.art-7232197b696746e2b1f47165fa735542 |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-03-11T14:52:50Z |
| publishDate | 2023-10-01 |
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| series | Frontiers in Plant Science |
| spelling | doaj.art-7232197b696746e2b1f47165fa7355422023-10-30T08:44:04ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2023-10-011410.3389/fpls.2023.12563381256338Paternal chromosome elimination of inducer triggers induction of double haploids in Brassica napusShihui Zhao0Shihui Zhao1Liangjun Huang2Liangjun Huang3Qing Zhang4Ying Zhou5Ying Zhou6Meicui Yang7Meicui Yang8Haoran Shi9Yun Li10Jin Yang11Chao Li12Xianhong Ge13Wanzhuo Gong14Jisheng Wang15Qiong Zou16Lanrong Tao17Zeming Kang18Zhuang Li19Chaowen Xiao20Qiong Hu21Shaohong Fu22Shaohong Fu23Chengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaAgricultural College, Sichuan Agricultural University, Chengdu, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaAgricultural College, Sichuan Agricultural University, Chengdu, ChinaCollege of Life Sciences, Sichuan University, Chengdu, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaAgricultural College, Sichuan Agricultural University, Chengdu, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaAgricultural College, Sichuan Agricultural University, Chengdu, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaOil Crops Research Institute, Chinese Academy of Agricultural Science, Wuhan, ChinaCollege of Plant Science and Technology of Huazhong Agricultural University, Wuhan, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaAgricultural College, Sichuan Agricultural University, Chengdu, ChinaCollege of Life Sciences, Sichuan University, Chengdu, ChinaOil Crops Research Institute, Chinese Academy of Agricultural Science, Wuhan, ChinaChengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, National Rapeseed Genetic Improvement Center, Chengdu, ChinaOil Crops Research Institute, Chinese Academy of Agricultural Science, Wuhan, ChinaA synthetic octoploid rapeseed, Y3380, induces maternal doubled haploids when used as a pollen donor to pollinate plant. However, the mechanism underlying doubled haploid formation remains elusive. We speculated that double haploid induction occurs as the inducer line’s chromosomes pass to the maternal egg cell, and the zygote is formed through fertilization. In the process of zygotic mitosis, the paternal chromosome is specifically eliminated. Part of the paternal gene might have infiltrated the maternal genome through homologous exchange during the elimination process. Then, the zygote haploid genome doubles (early haploid doubling, EH phenomenon), and the doubled zygote continues to develop into a complete embryo, finally forming doubled haploid offspring. To test our hypothesis, in the current study, the octoploid Y3380 line was back bred with the 4122-cp4-EPSPS exogenous gene used as a marker into hexaploid Y3380-cp4-EPSPS as paternal material to pollinate three different maternal materials. The fertilization process of crossing between the inducer line and the maternal parent was observed 48 h after pollination, and the fertilization rate reached 97.92% and 98.72%. After 12 d of pollination, the presence of cp4-EPSPS in the embryo was detected by in situ PCR, and at 13–23 d after pollination, the probability of F1 embryos containing cp4-EPSPS gene was up to 97.27%, but then declined gradually to 0% at 23–33 d. At the same time, the expression of cp4-EPSPS was observed by immunofluorescence in the 3rd to 29th day embryo. As the embryos developed, cp4-EPSPS marker genes were constantly lost, accompanied by embryonic death. After 30 d, the presence of cp4-EPSPS was not detected in surviving embryos. Meanwhile, SNP detection of induced offspring confirmed the existence of double haploids, further indicating that the induction process was caused by the loss of specificity of the paternal chromosome. The tetraploid-induced offspring showed infiltration of the induced line gene loci, with heterozygosity and homozygosity. Results indicated that the induced line chromosomes were eliminated during embryonic development, and the maternal haploid chromosomes were synchronously doubled in the embryo. These findings support our hypothesis and lay a theoretical foundation for further localization or cloning of functional genes involved in double haploid induction in rapeseed.https://www.frontiersin.org/articles/10.3389/fpls.2023.1256338/fullinducer linechromosome eliminationembryopaternal chromosomeBrassica napus L. |
| spellingShingle | Shihui Zhao Shihui Zhao Liangjun Huang Liangjun Huang Qing Zhang Ying Zhou Ying Zhou Meicui Yang Meicui Yang Haoran Shi Yun Li Jin Yang Chao Li Xianhong Ge Wanzhuo Gong Jisheng Wang Qiong Zou Lanrong Tao Zeming Kang Zhuang Li Chaowen Xiao Qiong Hu Shaohong Fu Shaohong Fu Paternal chromosome elimination of inducer triggers induction of double haploids in Brassica napus Frontiers in Plant Science inducer line chromosome elimination embryo paternal chromosome Brassica napus L. |
| title | Paternal chromosome elimination of inducer triggers induction of double haploids in Brassica napus |
| title_full | Paternal chromosome elimination of inducer triggers induction of double haploids in Brassica napus |
| title_fullStr | Paternal chromosome elimination of inducer triggers induction of double haploids in Brassica napus |
| title_full_unstemmed | Paternal chromosome elimination of inducer triggers induction of double haploids in Brassica napus |
| title_short | Paternal chromosome elimination of inducer triggers induction of double haploids in Brassica napus |
| title_sort | paternal chromosome elimination of inducer triggers induction of double haploids in brassica napus |
| topic | inducer line chromosome elimination embryo paternal chromosome Brassica napus L. |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2023.1256338/full |
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