An in vitro experiment was conducted to evaluate the effects of grain type on in vitro dry matter disappearance (IVDMD) and pH using ruminal fluid and a buffer reagent. Five beef cows were used for ruminal fluid colle...An in vitro experiment was conducted to evaluate the effects of grain type on in vitro dry matter disappearance (IVDMD) and pH using ruminal fluid and a buffer reagent. Five beef cows were used for ruminal fluid collection and ruminal fluid was pooled prior to use. The cows used for ruminal fluid collection were maintained on a forage-based diet (60:40 forage to concentrate) for 28 d prior to ruminal fluid collection. Three grain types were evaluated: 1) dry corn (89% dry matter;DRC), 2) high-moisture corn (72% dry matter;HMC) and 3) rye (89% dry matter;RYE). Data were analyzed as a completely randomized design. A total of twenty-one replications of each grain type were used for statistical analysis (n = 63 tubes total). IVDMD was greater (P ≤ 0.05) by 97.1% and 46.4% for RYE compared to DRC and HMC, respectively. Additionally, IVDMD was greater (P = 0.05) by 34.6% for HMC compared to DRC. Final pH was decreased (P ≤ 0.05) for RYE by 12.3% and 2.8% for RYE compared to DRC and HMC, respectively. Also, final pH was decreased (P = 0.05) by 9.8% for HMC compared to DRC. These data indicate that corn harvest method (dry vs. high-moisture) influences IVDMD and final pH and that rye has a greater disappearance of dry matter and lower final pH than corn under in vitro conditions.展开更多
Powdery mildew,caused by Blumeria graminis f.sp.tritici(Bgt),is a devastating disease that seriously threatens wheat yield and quality.To control this disease,host resistance is the most effective measure.Compared wit...Powdery mildew,caused by Blumeria graminis f.sp.tritici(Bgt),is a devastating disease that seriously threatens wheat yield and quality.To control this disease,host resistance is the most effective measure.Compared with the resistance genes from common wheat,alien resistance genes can better withstand infection of this highly variable pathogen.Development of elite alien germplasm resources with powdery mildew resistance and other key breeding traits is an attractive strategy in wheat breeding.In this study,three wheat-rye germplasm lines YT4-1,YT4-2,and YT4-3 were developed through hybridization between octoploid triticale and common wheat,out of which the lines YT4-1 and YT4-2 conferred adult-plant resistance(APR)to powdery mildew while the line YT4-3 was susceptible to powdery mildew during all of its growth stages.Using genomic in situ hybridization,multi-color fluorescence in situ hybridization,multi-color GISH,and molecular marker analysis,YT4-1,YT4-2,and YT4-3 were shown to be cytogenetically stable wheat-rye 6R addition and T1RS.1BL translocation line,6RL ditelosomic addition and T1RS.1BL translocation line,and T1RS.1BL translocation line,respectively.Compared with previously reported wheat-rye derivative lines carrying chromosome 6R,YT4-1 and YT4-2 showed stable APR without undesirable pleiotropic effects on agronomic traits.Therefore,these novel wheat-rye 6R derivative lines are expected to be promising bridge resources in wheat disease breeding.展开更多
Rye(Secale cereale L.)has been widely used to improve wheat(Triticum aestivum L.)cultivars.Oligo probes combined with non-denaturing fluorescence in situ hybridization(ND-FISH)technology provide a convenient and effic...Rye(Secale cereale L.)has been widely used to improve wheat(Triticum aestivum L.)cultivars.Oligo probes combined with non-denaturing fluorescence in situ hybridization(ND-FISH)technology provide a convenient and efficient way to identify individual rye chromosomes.However,suitable ND-FISH-positive oligo probes for recognizing specific segments of rye chromosomes are lacking.Five new ND-FISH-positive oligo probes:Oligo-5BL.46,Oligo-5A8080,Oligo-5A8080.1,Oligo-1AL.73,and Oligo-0R3,combined with two previously reported oligo probes,Oligo-44 and Oligo-45,were used in this study.Probes Oligo-5BL.46,Oligo-5A8080 and Oligo-44 produced signals only in intercalary regions of arms 1RS,5RS,and 5RL,respectively.Probe Oligo-5A8080.1 combined with probe Oligo-45 distinguished the intercalary regions of arms 1RS,5RS,and 6RS simultaneously.Oligo-5A8080 and Oligo-5A8080.1 revealed variation in the distribution of 5S rDNA sequences and polymorphism among 5R chromosomes.Probe Oligo-1AL.73 produced signals only on chromosomes 4R and 7R and contributed to the construction of an improved FISH map of chromosome 4RKu and to the confirmation of 4RLKu breakpoints in wheat-rye 4RLKu translocation chromosomes.Oligo-0R3 produced signals in the telomeric and subtelomeric regions of 14 rye chromosomes.These oligo probes also revealed five new tandem repeats in rye.Using the oligo probes reported in this study,the short arms of 1R,5R,and 6R and the long arms of 4R and 7R can be easily discriminated when these chromosomes are broken.展开更多
Rye(Secale cereale L., 2n=2x=14, RR) is a significant genetic resource for improving common wheat because of its resistance to multiple diseases and abiotic-stress tolerant traits. The 1RS chromosome from the German c...Rye(Secale cereale L., 2n=2x=14, RR) is a significant genetic resource for improving common wheat because of its resistance to multiple diseases and abiotic-stress tolerant traits. The 1RS chromosome from the German cultivated rye variety Petkus is critical in wheat breeding. However, its weakened disease resistance highlights the need to identify new resources. In the present study, a novel derived line called D27 was developed from common wheat and Mexico Rye.Cytological observations characterized the karyotype of D27 as 2n=42=21 Ⅱ. Genomic in situ hybridization indicated that a pair of whole-arm translocated Mexico Rye chromosomes were inherited typically in the mitotic and meiosis stages of D27. Experiments using fluorescence in situ hybridization(FISH) and gliadin electrophoresis showed that D27 lacked wheat 1DS chromosomes. They were replaced by 1RS chromosomes of Mexico Rye, supported by wheat simple-sequence repeat markers, rye sequence characterized amplified region markers, and wheat 40K SNP array analysis.The wheat 1DS chromosomes could not be detected by molecular markers and wheat SNP array, but the presence of rye 1RS chromosomes was confirmed. Agronomic trait assessments indicated that D27 had a higher tiller number and enhanced stripe rust and powdery mildew resistance. In addition, dough properties analysis showed that replacing 1DS led to higher viscosity and lower dough elasticity in D27, which was beneficial for cake making. In conclusion, the novel cytogenetically stable common wheat–Mexico Rye T1DL·1RS translocation line D27 offers excellent potential as outstanding germplasm in wheat breeding programs focusing on disease resistance and yield improvement. Additionally,it can be valuable for researching the rye 1RS chromosome’s genetic diversity.展开更多
Rye(Secale cereale)is a valuable gene donor for wheat improvement,especially for its resistance to diseases.Developing rye-derived resistance sources is important for wheat breeding.In the present study,two wheat-rye ...Rye(Secale cereale)is a valuable gene donor for wheat improvement,especially for its resistance to diseases.Developing rye-derived resistance sources is important for wheat breeding.In the present study,two wheat-rye derivatives,designated JS016 and JS110,were produced by crossing common wheat cultivar Yangmai 23 with Pakistani rye accession W2A.Using sequential genomic in situ hybridization(GISH)and multicolor fluorescence in situ hybridization(mc-FISH),JS016 and JS110 were identified as a T6BS.6RL translocation line and a T6BS.6BL6RL translocation line,respectively.Ten newly 6RL chromosome arm-specific markers were developed and used to confirm the 6RL translocation.The wheat 55K single-nucleotide polymorphism(SNP)array further verified the molecular cytogenetic identification results above and clarified their breakpoints at 430.9 and 523.0 Mb of chromosome 6B in JS016 and JS110,respectively.Resistance spectrum and allelism test demonstrated that JS016 and JS110 possessed novel powdery mildew resistance gene(s)that was derived from the 6RL translocation but differed from Pm20.Moreover,JS016 and JS110 had better agronomic traits than the previously reported 6RL translocation line carrying Pm20.To efficiently transfer and detect the 6RL translocation from JS016 and JS110,one 6RL-specific Kompetitive allele specific PCR(KASP)marker was developed and validated in high throughput marker-assisted selection(MAS).展开更多
Species closely related to wheat are important genetic resources for agricultural production,functional genomics studies and wheat improvement.In this study,a wheat gene related to regeneration,TaWOX5,was applied to e...Species closely related to wheat are important genetic resources for agricultural production,functional genomics studies and wheat improvement.In this study,a wheat gene related to regeneration,TaWOX5,was applied to establish the Agrobacterium-mediated transformation systems of Triticum monococcum,hexaploid triticale,and rye(Secale cereale L.)using their immature embryos.Transgenic plants were efficiently generated.During the transformation process,the Agrobacterium infection efficiency was assessed by histochemical staining forβ-glucuronidase(GUS).Finally,the transgenic nature of regenerated plants was verified by polymerase chain reaction(PCR)-based genotyping for the presence of the GUS and bialaphos resistance(bar)genes,histochemical staining for GUS protein,and the QuickStix strip assay for bar protein.The transformation efficiency of T.monococcum genotype PI428182 was 94.4%;the efficiencies of four hexaploid triticale genotypes Lin456,ZS3297,ZS1257,and ZS3224 were 52.1,41.2,19.4,and 16.0%,respectively;and the transformation efficiency of rye cultivar Lanzhou Heimai was 7.8%.Fluorescence in situ hybridization(FISH)and genomic in situ hybridization(GISH)analyses indicated that the GUS transgenes were integrated into the distal or near centromere(proximal)regions of the chromosomes in transgenic T.monococcum and hexaploid triticale plants.In the transgenic hexaploid triticale plants,the foreign DNA fragment was randomly integrated into the AABB and RR genomes.Furthermore,the transgene was almost stably inherited in the next generation by Mendel’s law.The findings in this study will promote the genetic improvement of the three plant species for grain or forage production and the improvement of cereal species including wheat for functional genomics studies.展开更多
文摘An in vitro experiment was conducted to evaluate the effects of grain type on in vitro dry matter disappearance (IVDMD) and pH using ruminal fluid and a buffer reagent. Five beef cows were used for ruminal fluid collection and ruminal fluid was pooled prior to use. The cows used for ruminal fluid collection were maintained on a forage-based diet (60:40 forage to concentrate) for 28 d prior to ruminal fluid collection. Three grain types were evaluated: 1) dry corn (89% dry matter;DRC), 2) high-moisture corn (72% dry matter;HMC) and 3) rye (89% dry matter;RYE). Data were analyzed as a completely randomized design. A total of twenty-one replications of each grain type were used for statistical analysis (n = 63 tubes total). IVDMD was greater (P ≤ 0.05) by 97.1% and 46.4% for RYE compared to DRC and HMC, respectively. Additionally, IVDMD was greater (P = 0.05) by 34.6% for HMC compared to DRC. Final pH was decreased (P ≤ 0.05) for RYE by 12.3% and 2.8% for RYE compared to DRC and HMC, respectively. Also, final pH was decreased (P = 0.05) by 9.8% for HMC compared to DRC. These data indicate that corn harvest method (dry vs. high-moisture) influences IVDMD and final pH and that rye has a greater disappearance of dry matter and lower final pH than corn under in vitro conditions.
基金This research was supported by the National Key Research and Development Program of China(2021YFD1200600)the National Natural Science Foundation of China(32272105).
文摘Powdery mildew,caused by Blumeria graminis f.sp.tritici(Bgt),is a devastating disease that seriously threatens wheat yield and quality.To control this disease,host resistance is the most effective measure.Compared with the resistance genes from common wheat,alien resistance genes can better withstand infection of this highly variable pathogen.Development of elite alien germplasm resources with powdery mildew resistance and other key breeding traits is an attractive strategy in wheat breeding.In this study,three wheat-rye germplasm lines YT4-1,YT4-2,and YT4-3 were developed through hybridization between octoploid triticale and common wheat,out of which the lines YT4-1 and YT4-2 conferred adult-plant resistance(APR)to powdery mildew while the line YT4-3 was susceptible to powdery mildew during all of its growth stages.Using genomic in situ hybridization,multi-color fluorescence in situ hybridization,multi-color GISH,and molecular marker analysis,YT4-1,YT4-2,and YT4-3 were shown to be cytogenetically stable wheat-rye 6R addition and T1RS.1BL translocation line,6RL ditelosomic addition and T1RS.1BL translocation line,and T1RS.1BL translocation line,respectively.Compared with previously reported wheat-rye derivative lines carrying chromosome 6R,YT4-1 and YT4-2 showed stable APR without undesirable pleiotropic effects on agronomic traits.Therefore,these novel wheat-rye 6R derivative lines are expected to be promising bridge resources in wheat disease breeding.
基金the National Key Research and Development Program of China(2016YFD0102001)the National Natural Science Foundation of China(31770373).
文摘Rye(Secale cereale L.)has been widely used to improve wheat(Triticum aestivum L.)cultivars.Oligo probes combined with non-denaturing fluorescence in situ hybridization(ND-FISH)technology provide a convenient and efficient way to identify individual rye chromosomes.However,suitable ND-FISH-positive oligo probes for recognizing specific segments of rye chromosomes are lacking.Five new ND-FISH-positive oligo probes:Oligo-5BL.46,Oligo-5A8080,Oligo-5A8080.1,Oligo-1AL.73,and Oligo-0R3,combined with two previously reported oligo probes,Oligo-44 and Oligo-45,were used in this study.Probes Oligo-5BL.46,Oligo-5A8080 and Oligo-44 produced signals only in intercalary regions of arms 1RS,5RS,and 5RL,respectively.Probe Oligo-5A8080.1 combined with probe Oligo-45 distinguished the intercalary regions of arms 1RS,5RS,and 6RS simultaneously.Oligo-5A8080 and Oligo-5A8080.1 revealed variation in the distribution of 5S rDNA sequences and polymorphism among 5R chromosomes.Probe Oligo-1AL.73 produced signals only on chromosomes 4R and 7R and contributed to the construction of an improved FISH map of chromosome 4RKu and to the confirmation of 4RLKu breakpoints in wheat-rye 4RLKu translocation chromosomes.Oligo-0R3 produced signals in the telomeric and subtelomeric regions of 14 rye chromosomes.These oligo probes also revealed five new tandem repeats in rye.Using the oligo probes reported in this study,the short arms of 1R,5R,and 6R and the long arms of 4R and 7R can be easily discriminated when these chromosomes are broken.
基金supported by the National Natural Science Foundation of China (31771785)the Key Research and Development Program of Shaanxi, China (2018ZDXM-NY-006)。
文摘Rye(Secale cereale L., 2n=2x=14, RR) is a significant genetic resource for improving common wheat because of its resistance to multiple diseases and abiotic-stress tolerant traits. The 1RS chromosome from the German cultivated rye variety Petkus is critical in wheat breeding. However, its weakened disease resistance highlights the need to identify new resources. In the present study, a novel derived line called D27 was developed from common wheat and Mexico Rye.Cytological observations characterized the karyotype of D27 as 2n=42=21 Ⅱ. Genomic in situ hybridization indicated that a pair of whole-arm translocated Mexico Rye chromosomes were inherited typically in the mitotic and meiosis stages of D27. Experiments using fluorescence in situ hybridization(FISH) and gliadin electrophoresis showed that D27 lacked wheat 1DS chromosomes. They were replaced by 1RS chromosomes of Mexico Rye, supported by wheat simple-sequence repeat markers, rye sequence characterized amplified region markers, and wheat 40K SNP array analysis.The wheat 1DS chromosomes could not be detected by molecular markers and wheat SNP array, but the presence of rye 1RS chromosomes was confirmed. Agronomic trait assessments indicated that D27 had a higher tiller number and enhanced stripe rust and powdery mildew resistance. In addition, dough properties analysis showed that replacing 1DS led to higher viscosity and lower dough elasticity in D27, which was beneficial for cake making. In conclusion, the novel cytogenetically stable common wheat–Mexico Rye T1DL·1RS translocation line D27 offers excellent potential as outstanding germplasm in wheat breeding programs focusing on disease resistance and yield improvement. Additionally,it can be valuable for researching the rye 1RS chromosome’s genetic diversity.
基金supported by the National Natural Science Foundation of China(32171990 and 32072053)Key Research and Development Program of Zhenjiang(NY2021001)+4 种基金State Key Laboratory of Plant Cell and Chromosome Engineering(PCCE-KF-2021-05 and PCCE-KF-2022-07)State Key Laboratory of Crop Biology in Shandong Agricultural University(2021KF01)Natural Science Foundation of the Jiangsu Higher Education institutions of China(21KJB210004)Open Project Funding of State Key Laboratory of Crop Stress Adaptation and Improvement(CX1130A0920014)Key Research and Development Program of Shandong Province(2020CXGC010805).
文摘Rye(Secale cereale)is a valuable gene donor for wheat improvement,especially for its resistance to diseases.Developing rye-derived resistance sources is important for wheat breeding.In the present study,two wheat-rye derivatives,designated JS016 and JS110,were produced by crossing common wheat cultivar Yangmai 23 with Pakistani rye accession W2A.Using sequential genomic in situ hybridization(GISH)and multicolor fluorescence in situ hybridization(mc-FISH),JS016 and JS110 were identified as a T6BS.6RL translocation line and a T6BS.6BL6RL translocation line,respectively.Ten newly 6RL chromosome arm-specific markers were developed and used to confirm the 6RL translocation.The wheat 55K single-nucleotide polymorphism(SNP)array further verified the molecular cytogenetic identification results above and clarified their breakpoints at 430.9 and 523.0 Mb of chromosome 6B in JS016 and JS110,respectively.Resistance spectrum and allelism test demonstrated that JS016 and JS110 possessed novel powdery mildew resistance gene(s)that was derived from the 6RL translocation but differed from Pm20.Moreover,JS016 and JS110 had better agronomic traits than the previously reported 6RL translocation line carrying Pm20.To efficiently transfer and detect the 6RL translocation from JS016 and JS110,one 6RL-specific Kompetitive allele specific PCR(KASP)marker was developed and validated in high throughput marker-assisted selection(MAS).
基金supported by grants from the National Natural Science Foundation of China (31971946)the Technology Innovation Program of the Chinese Academy of Agricultural Sciences, China (2060302-2-23 and ASTIP-2060302-2-19).
文摘Species closely related to wheat are important genetic resources for agricultural production,functional genomics studies and wheat improvement.In this study,a wheat gene related to regeneration,TaWOX5,was applied to establish the Agrobacterium-mediated transformation systems of Triticum monococcum,hexaploid triticale,and rye(Secale cereale L.)using their immature embryos.Transgenic plants were efficiently generated.During the transformation process,the Agrobacterium infection efficiency was assessed by histochemical staining forβ-glucuronidase(GUS).Finally,the transgenic nature of regenerated plants was verified by polymerase chain reaction(PCR)-based genotyping for the presence of the GUS and bialaphos resistance(bar)genes,histochemical staining for GUS protein,and the QuickStix strip assay for bar protein.The transformation efficiency of T.monococcum genotype PI428182 was 94.4%;the efficiencies of four hexaploid triticale genotypes Lin456,ZS3297,ZS1257,and ZS3224 were 52.1,41.2,19.4,and 16.0%,respectively;and the transformation efficiency of rye cultivar Lanzhou Heimai was 7.8%.Fluorescence in situ hybridization(FISH)and genomic in situ hybridization(GISH)analyses indicated that the GUS transgenes were integrated into the distal or near centromere(proximal)regions of the chromosomes in transgenic T.monococcum and hexaploid triticale plants.In the transgenic hexaploid triticale plants,the foreign DNA fragment was randomly integrated into the AABB and RR genomes.Furthermore,the transgene was almost stably inherited in the next generation by Mendel’s law.The findings in this study will promote the genetic improvement of the three plant species for grain or forage production and the improvement of cereal species including wheat for functional genomics studies.