Both cDNA and a genomic DNA fragment encoding a new potato proteinase inhibitor Ⅱ were isolated from a diploid potato IVP101 （Solanum phurejia L.） and named PINII-2x. Nucleotlde sequencing confirmed that the DNA fr...Both cDNA and a genomic DNA fragment encoding a new potato proteinase inhibitor Ⅱ were isolated from a diploid potato IVP101 （Solanum phurejia L.） and named PINII-2x. Nucleotlde sequencing confirmed that the DNA fragment of PINll-2xwas 580 bp, including a 115-bp intron and two exons. The deduced PINII-2x proteln contained an Intact signal peptide and two active sites. The PINII-2x gene and its deduced PINII-2x protein had 88% and 93% homology with another tetraploid potato proteinase inhibitor Ⅱ, respectively. Northern blotting analysis Indicated that the mRNA of PINII-2x gene was wound induced in potato leaves. Binary vector pNAR301 and pNAR302 were constructed for rice transformation, in which the PINII-2x cDNA was driven, respectively, by rice actin I promoter （Actl） and maize ubiquitin promoter （Ubll）. Via an Agrobacteriummediated method, these two constructs were transferred into japonica rice cv. Xiushui 63. PCR and Southern blotUng analysis for transgenic rice revealed the integration of the PINII-2x gene. Northern blotting analysis also confirmed transcripts of the PINII.2x gene in transgenic rice plants. Insect bloassays using stripe stem borer （Chilo auppressalis Walker） demonstrated that the average weight and body length of larvae In transgenic plants were only nearly 50% and 61% of those of larvae in control plants, respectively. These results Indicate that the PINII-2x gene should be an effective insect-resistance gene and could be valuable for application in crop breeding for insect resistance.展开更多
In potato breeding,it is difficult to improve the traits of interest at the tetraploid level due to the tetrasomic inheritance.A promising alternative is diploid breeding.Thus it is necessary to assess the genetic div...In potato breeding,it is difficult to improve the traits of interest at the tetraploid level due to the tetrasomic inheritance.A promising alternative is diploid breeding.Thus it is necessary to assess the genetic diversity of diploid potato germplasm for efficient exploration and deployment of desirable traits.In this study,we used SSR markers to evaluate the genetic diversity of diploid potato cultivars.To screen polymorphic SSR markers,55 pairs of SSR primers were employed to amplify 39 cultivars with relatively distant genetic relationships.Among them,12 SSR markers with high polymorphism located at 12 chromosomes were chosen to evaluate the genetic diversity of 192 diploid potato cultivars.The primers produced 6 to 18 bands with an average of 8.2 bands per primer.In total,98 bands were amplified from 192 cultivars,and 97 of them were polymorphic.Cluster analysis using UPGMA showed the genetic relationships of all accessions tested:186 of the 192 accessions could be distinguished by only 12 pairs of SSR primers,and the 192 diploid cultivars were divided into 11 groups,and 83.3% constituted the first group.Clustering results showed relatively low genetic diversity among 192 diploid cultivars,with closer relationship at the molecular level.The results can provide molecular basis for diploid potato breeding.展开更多
Reinventing the tetraploid potato into a seed-propagated,diploid,hybrid potato would significantly accelerate potato breeding.In this regard,the development of highly homozygous inbred lines is a prerequisite for bree...Reinventing the tetraploid potato into a seed-propagated,diploid,hybrid potato would significantly accelerate potato breeding.In this regard,the development of highly homozygous inbred lines is a prerequisite for breeding hybrid potatoes,but self-incompatibility and inbreeding depression present challenges for developing pure inbred lines.To resolve this impediment,we developed a doubled haploid(DH)technology,based on mutagenesis of the potato DOMAIN OF UNKNOWN FUNCTION 679 membrane protein(StDMP)gene.Here,we show that a deficiency in StDMP allows the generation of maternal haploids for generating diploid potato lines.An exercisable protocol,involving hybridization,fluorescent marker screening,molecular and flow cytometric identification,and doubling with colchicine generates nearly 100%homozygous diploid potato lines.This dmp-triggered haploid induction(HI)system greatly shortens the breeding process and offers a robust method for generating diploid potato inbred lines with high purity.展开更多
The narrow genetic base in potato (Solanum tuberosum L.) limits the progress in cultivar development.The rich diploid germplasm in the origin center of potato provide a unique resource for improvement of tetraploid po...The narrow genetic base in potato (Solanum tuberosum L.) limits the progress in cultivar development.The rich diploid germplasm in the origin center of potato provide a unique resource for improvement of tetraploid potatoes.Seven newly developed diploid hybrids with 2n pollen production,all of which have S. phureja background,were developed and evaluated for their value in potato breeding.They were crossed as male parnets to six tetraploid Solanum tuberosum cultivars,and seeds in large quantity from eleven crosses were obtained.Main agronomic traits,such as tuber yield,tuber number,mean tuber weight,tuber shape,eye depth,skin smoothness,flesh color,and specific gravity,were measured for 4x 2x tetraploid progenies in seedling generation,and their parents as well.All of the diploid hybrids had some merit for specific traits and the DH39 was more promising;high specific gravity trait in some diploid hybrids was successfully introgressed into tetroploid progenies via 4x 2x crosses.These diploid hybrids have potential value in potato breeding.展开更多
基金Supported by the Project of Jiangsu Province(BG2001305)the Program for Changjiang Scholars and Innovative Research Team in University
文摘Both cDNA and a genomic DNA fragment encoding a new potato proteinase inhibitor Ⅱ were isolated from a diploid potato IVP101 （Solanum phurejia L.） and named PINII-2x. Nucleotlde sequencing confirmed that the DNA fragment of PINll-2xwas 580 bp, including a 115-bp intron and two exons. The deduced PINII-2x proteln contained an Intact signal peptide and two active sites. The PINII-2x gene and its deduced PINII-2x protein had 88% and 93% homology with another tetraploid potato proteinase inhibitor Ⅱ, respectively. Northern blotting analysis Indicated that the mRNA of PINII-2x gene was wound induced in potato leaves. Binary vector pNAR301 and pNAR302 were constructed for rice transformation, in which the PINII-2x cDNA was driven, respectively, by rice actin I promoter （Actl） and maize ubiquitin promoter （Ubll）. Via an Agrobacteriummediated method, these two constructs were transferred into japonica rice cv. Xiushui 63. PCR and Southern blotUng analysis for transgenic rice revealed the integration of the PINII-2x gene. Northern blotting analysis also confirmed transcripts of the PINII.2x gene in transgenic rice plants. Insect bloassays using stripe stem borer （Chilo auppressalis Walker） demonstrated that the average weight and body length of larvae In transgenic plants were only nearly 50% and 61% of those of larvae in control plants, respectively. These results Indicate that the PINII-2x gene should be an effective insect-resistance gene and could be valuable for application in crop breeding for insect resistance.
基金funded by the National Natural Science Foundation of China(NSFC:31201257)the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(CAAS-ASTIP-AGISCAAS)
文摘In potato breeding,it is difficult to improve the traits of interest at the tetraploid level due to the tetrasomic inheritance.A promising alternative is diploid breeding.Thus it is necessary to assess the genetic diversity of diploid potato germplasm for efficient exploration and deployment of desirable traits.In this study,we used SSR markers to evaluate the genetic diversity of diploid potato cultivars.To screen polymorphic SSR markers,55 pairs of SSR primers were employed to amplify 39 cultivars with relatively distant genetic relationships.Among them,12 SSR markers with high polymorphism located at 12 chromosomes were chosen to evaluate the genetic diversity of 192 diploid potato cultivars.The primers produced 6 to 18 bands with an average of 8.2 bands per primer.In total,98 bands were amplified from 192 cultivars,and 97 of them were polymorphic.Cluster analysis using UPGMA showed the genetic relationships of all accessions tested:186 of the 192 accessions could be distinguished by only 12 pairs of SSR primers,and the 192 diploid cultivars were divided into 11 groups,and 83.3% constituted the first group.Clustering results showed relatively low genetic diversity among 192 diploid cultivars,with closer relationship at the molecular level.The results can provide molecular basis for diploid potato breeding.
基金supported by the Guangdong Major Project of Basic and Applied Basic Research(2021B0301030004)the National Natural Science Foundation of China(Grant No.31991185,31902027)supported by the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Science.Author information。
文摘Reinventing the tetraploid potato into a seed-propagated,diploid,hybrid potato would significantly accelerate potato breeding.In this regard,the development of highly homozygous inbred lines is a prerequisite for breeding hybrid potatoes,but self-incompatibility and inbreeding depression present challenges for developing pure inbred lines.To resolve this impediment,we developed a doubled haploid(DH)technology,based on mutagenesis of the potato DOMAIN OF UNKNOWN FUNCTION 679 membrane protein(StDMP)gene.Here,we show that a deficiency in StDMP allows the generation of maternal haploids for generating diploid potato lines.An exercisable protocol,involving hybridization,fluorescent marker screening,molecular and flow cytometric identification,and doubling with colchicine generates nearly 100%homozygous diploid potato lines.This dmp-triggered haploid induction(HI)system greatly shortens the breeding process and offers a robust method for generating diploid potato inbred lines with high purity.
文摘The narrow genetic base in potato (Solanum tuberosum L.) limits the progress in cultivar development.The rich diploid germplasm in the origin center of potato provide a unique resource for improvement of tetraploid potatoes.Seven newly developed diploid hybrids with 2n pollen production,all of which have S. phureja background,were developed and evaluated for their value in potato breeding.They were crossed as male parnets to six tetraploid Solanum tuberosum cultivars,and seeds in large quantity from eleven crosses were obtained.Main agronomic traits,such as tuber yield,tuber number,mean tuber weight,tuber shape,eye depth,skin smoothness,flesh color,and specific gravity,were measured for 4x 2x tetraploid progenies in seedling generation,and their parents as well.All of the diploid hybrids had some merit for specific traits and the DH39 was more promising;high specific gravity trait in some diploid hybrids was successfully introgressed into tetroploid progenies via 4x 2x crosses.These diploid hybrids have potential value in potato breeding.
