Beany flavor induced by three lipoxygenases(LOXs, including LOX1, LOX2, and LOX3)restricts human consumption of soybean. It is desirable to generate lipoxygenase-free new mutant lines to improve the eating quality of ...Beany flavor induced by three lipoxygenases(LOXs, including LOX1, LOX2, and LOX3)restricts human consumption of soybean. It is desirable to generate lipoxygenase-free new mutant lines to improve the eating quality of soybean oil and protein products. In this study, a pooled clustered regularly interspaced short palindromic repeats(CRISPR)-CRISPRassociated protein 9(Cas9) strategy targeting three GmLox genes(GmLox1, GmLox2, and GmLox3) was applied and 60 T_0 positive transgenic plants were generated, carrying combinations of sg RNAs and mutations. Among them, GmLox-28 and GmLox-60 were gmlox1 gmlox2 gmlox3 triple mutants and GmLox-40 was a gmlox1 gmlox2 double mutant.Sequencing of T_1 mutant plants derived from GmLox-28, GmLox-60, and GmLox-40 showed that mutation in the GmLox gene was inherited by the next generation. Colorimetric assay revealed that plants carrying different combinations of mutations lost the corresponding lipoxygenase activities. Transgene-free mutants were obtained by screening the T_2 generation of lipoxygenase-free mutant lines(GmLox-28 and GmLox-60). These transgeneand lipoxygenase-free mutants could be used for soybean beany flavor reduction without restriction by regulatory frameworks governing transgenic organisms.展开更多
The level of genetic variation within a breeding population affects the effectiveness of selection strategies for genetic improvement.The relationship between genetic variation level within Pinus tabuliformis breeding...The level of genetic variation within a breeding population affects the effectiveness of selection strategies for genetic improvement.The relationship between genetic variation level within Pinus tabuliformis breeding populations and selection strategies or selection effectiveness is not fully investigated.Here,we compared the selection effectiveness of combined and individual direct selection strategies using half-and full-sib families produced from advanced-generation P.tabuliformis seed orchard as our test populations.Our results revealed that,within half-sib families,average diameter at breast height(DBH),tree height,and volume growth of superior individuals selected by the direct selection strategy were higher by 7.72%,18.56%,and 31.01%,respectively,than those selected by the combined selection strategy.Furthermore,significant differences(P<0.01)were observed between the two strategies in terms of the expected genetic gains for average tree height and volume.In contrast,within full-sib families,the differences in tree average DBH,height,and volume between the two selection strategies were relatively minor with increase of 0.17%,2.73%,and 2.21%,respectively,and no significant differences were found in the average expected genetic gains for the studied traits.Half-sib families exhibited greater phenotypic and genetic variation,resulting in improved selection efficiency with the direct selection strategy but also introduced a level of inbreeding risk.Based on genetic distance estimates using molecular markers,our comparative seed orchard design analysis showed that the Improved Adaptive Genetic Programming Algorithm(IAPGA)reduced the average inbreeding coefficient by 14.36% and 14.73% compared to sequential and random designs,respectively.In conclusion,the combination of the direct selection strategy with IAPGA seed orchard design aimed at minimizing inbreeding offered an efficient approach for establishing advanced-generation P.tabuliformis seed orchards.展开更多
基金supported by funds from the National Key Research and Development Program of China(2016YFD0100700)to Y.G。
文摘Beany flavor induced by three lipoxygenases(LOXs, including LOX1, LOX2, and LOX3)restricts human consumption of soybean. It is desirable to generate lipoxygenase-free new mutant lines to improve the eating quality of soybean oil and protein products. In this study, a pooled clustered regularly interspaced short palindromic repeats(CRISPR)-CRISPRassociated protein 9(Cas9) strategy targeting three GmLox genes(GmLox1, GmLox2, and GmLox3) was applied and 60 T_0 positive transgenic plants were generated, carrying combinations of sg RNAs and mutations. Among them, GmLox-28 and GmLox-60 were gmlox1 gmlox2 gmlox3 triple mutants and GmLox-40 was a gmlox1 gmlox2 double mutant.Sequencing of T_1 mutant plants derived from GmLox-28, GmLox-60, and GmLox-40 showed that mutation in the GmLox gene was inherited by the next generation. Colorimetric assay revealed that plants carrying different combinations of mutations lost the corresponding lipoxygenase activities. Transgene-free mutants were obtained by screening the T_2 generation of lipoxygenase-free mutant lines(GmLox-28 and GmLox-60). These transgeneand lipoxygenase-free mutants could be used for soybean beany flavor reduction without restriction by regulatory frameworks governing transgenic organisms.
基金financially supported by the Biological BreedingNational Science and Technology Major Project(2023ZD0405806)the National Key R&D Program for the 14th Five-Year Plan in China(2022YFD2200304).
文摘The level of genetic variation within a breeding population affects the effectiveness of selection strategies for genetic improvement.The relationship between genetic variation level within Pinus tabuliformis breeding populations and selection strategies or selection effectiveness is not fully investigated.Here,we compared the selection effectiveness of combined and individual direct selection strategies using half-and full-sib families produced from advanced-generation P.tabuliformis seed orchard as our test populations.Our results revealed that,within half-sib families,average diameter at breast height(DBH),tree height,and volume growth of superior individuals selected by the direct selection strategy were higher by 7.72%,18.56%,and 31.01%,respectively,than those selected by the combined selection strategy.Furthermore,significant differences(P<0.01)were observed between the two strategies in terms of the expected genetic gains for average tree height and volume.In contrast,within full-sib families,the differences in tree average DBH,height,and volume between the two selection strategies were relatively minor with increase of 0.17%,2.73%,and 2.21%,respectively,and no significant differences were found in the average expected genetic gains for the studied traits.Half-sib families exhibited greater phenotypic and genetic variation,resulting in improved selection efficiency with the direct selection strategy but also introduced a level of inbreeding risk.Based on genetic distance estimates using molecular markers,our comparative seed orchard design analysis showed that the Improved Adaptive Genetic Programming Algorithm(IAPGA)reduced the average inbreeding coefficient by 14.36% and 14.73% compared to sequential and random designs,respectively.In conclusion,the combination of the direct selection strategy with IAPGA seed orchard design aimed at minimizing inbreeding offered an efficient approach for establishing advanced-generation P.tabuliformis seed orchards.
