Phosphite (Phi)-based fungicides are used to control the oomycete Phytophthora infestans which causes late blight disease, the most devastating disease in potatoes. In order to examine the effects of Phi-based fungici...Phosphite (Phi)-based fungicides are used to control the oomycete Phytophthora infestans which causes late blight disease, the most devastating disease in potatoes. In order to examine the effects of Phi-based fungicides on potato tubers through foliar or post-harvest application, a metabolite profiling approach based on gas chromatography coupled to mass spectrometry (GC-MS) has been established. A total of 132 metabolites were detected using the GC-MS approach. Among these, 34 metabolites were identified after normalization and annotated with a compound name with standard mass spectral library. Metabolomic analysis of Phi-treated plants showed significant differences in the levels of many metabolites especially amino acids. Multivariate statistical approaches, such as principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), were employed to explore the relationships between metabolites to detect group differences. A good discrimination between the control and the Phi-treated plants was observed, which demonstrated that significant changes in the metabolite profile have been caused by the two different Phi applications (foliar or post-harvest). This finding suggests that the alteration of specific metabolite levels by accumulation of Phi can lead to resistance against the pathogen.展开更多
An efficient and rapid Agrobacterium tumefaciens-mediated transformation protocol was developed to generate activation-tagged mutant lines with the aim of large-scale functional analysis of the potato genome. The expl...An efficient and rapid Agrobacterium tumefaciens-mediated transformation protocol was developed to generate activation-tagged mutant lines with the aim of large-scale functional analysis of the potato genome. The explants were inoculated with an Agrobacterium strain harboring the binary plasmid pSKI074 containing four CaMV 35S enhancers in the T-DNA region which activates the downstream genes in the host plant after its integration. Various parameters investigated to increase transformation efficiency were the type and age of explant, cultivar, hormone combinations, preculture of explants, period of co-cultivation with bacteria and concentration of bacterial cultures used for transformation. Stem explants from 5 week old plantlets of cv. Bintje which had undergone phytohormone pretreatment for 4 days, inoculation with diluted bacterial concentration of OD600 = 0.2 containing acetosyringone followed by 2 days of co-cultivation and selection in media with IAA and trans-zeatin all helped in greatly improving the transformation efficiency. The total time required from infection to rooted shoots was 6-7 weeks. Initial evidence for stable integration and expression of the transgenes by PCR analysis showed that over 93% of the regenerated lines were transgenic and this was confirmed by Southern hybridization.展开更多
Potato late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating diseases in the agricultural sector around the world. Many genes (R genes) conferring resistance to late blight have be...Potato late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating diseases in the agricultural sector around the world. Many genes (R genes) conferring resistance to late blight have been identified in various potato species and most of these R genes have been used in potato breeding. The aim of this study was to develop and validate PCR-based assays for the R genes Rpi-blb1, Rpi-blb2, Rpi-blb3 and Rpi-bt1, to distinguish between late blight resistant and late blight susceptible potato progeny in the given breeding background. A total of 100 breeding progeny were screened for the presence of these R genes and tested for resistance against P. infestans mating type A2, genotype US-8 strain, using detached leaf and tuber rot assays. PCR products for the Rpi-blb1 and Rpi-bt1 resistance genes were identified in the resistant progeny but were absent in the susceptible ones;therefore these PCR assays could differentiate between late blight resistant and susceptible plants. Genotypic data from the DNA markers derived from the Rpi-blb1 and Rpi-bt1 genes was found to correlate with the phenotypic data for foliar late blight but not with data for tuber rot. Our results demonstrate that markers derived from these two R genes could be useful for marker-assisted selection (MAS) for foliar late blight resistance in potato breeding programs.展开更多
Total phenolics, anthocyanins and antioxidant capacity of five coloured/pigmented (AR2009-10, Adirondack Red, Adirondack Blue, Congo, and POROIPG22-1) and two unpigmented potato genotypes (“Anuschka” and “Russet Bu...Total phenolics, anthocyanins and antioxidant capacity of five coloured/pigmented (AR2009-10, Adirondack Red, Adirondack Blue, Congo, and POROIPG22-1) and two unpigmented potato genotypes (“Anuschka” and “Russet Burbank”) were assessed in fresh (with and without skin) and commercially processed/cooked/reconstituted products. Ascorbate profiles of the seven genotypes also were investigated using fresh tuber/tissue only. The results showed that genotypes greatly varied in their contents of bioactive compounds. Ascorbate profiles of the genotypes were not associated to any particular flesh colour/pigment. However, the pigmented potatoes had 1.5 to 2.5 times more the phenolics, 2 to 3 times more antioxidant capacity and higher levels of anthocyanins (13.98 to 38.57 mg C3GE-100g FW) compared to unpigmented genotypes. No anthocyanins were detected in the unpigmented potatoes. Significant losses of total phenolics, anthocyanins and total antioxidant capacity were found during peeling (18% - 23%), blanching process (40% - 60%) and further cooking/ reconstitution (7% - 12%) with no prominent genotype differences. Together, 65 to 90% of these bioactive compounds were lost during processing. The results suggested that pigmented potatoes contained higher amounts of total phenolics and anthocyanins and blanching step took away the most of the original bioactive compounds.展开更多
In Brassicaceae, myrosinase catalyzes the hydrolysis of glucosinolate and plays an important role in anti-herbivore defense. We have cloned and characterized the full- length complementary DNA of myrosinase gene from ...In Brassicaceae, myrosinase catalyzes the hydrolysis of glucosinolate and plays an important role in anti-herbivore defense. We have cloned and characterized the full- length complementary DNA of myrosinase gene from Brassicaparachinensis that exhibits high sequence identity with myrosinase genes from other Brassica species. To investigate the role of this myrosinase in defense against the diamondback moth （Plutella xylostella）, we constructed an RNA-interference （RNAi） cassette expressing a double-stranded RNA that targeted myrosinase and transfected it into B. parachinensis. Myrosinase was sup- pressed in the resulting transgenic plants. Diamondback moth larvae feeding on transgenic plants had lower larval and pupal weights, longer pupal duration, and lower fecundity than those feeding on non-transgenic plants, suggesting that the diamondback moth has adapted to the glucosinolate-myrosinase defensive system. Therefore, the suppression of myrosinase is a potential approach for controlling the diamondback moth.展开更多
文摘Phosphite (Phi)-based fungicides are used to control the oomycete Phytophthora infestans which causes late blight disease, the most devastating disease in potatoes. In order to examine the effects of Phi-based fungicides on potato tubers through foliar or post-harvest application, a metabolite profiling approach based on gas chromatography coupled to mass spectrometry (GC-MS) has been established. A total of 132 metabolites were detected using the GC-MS approach. Among these, 34 metabolites were identified after normalization and annotated with a compound name with standard mass spectral library. Metabolomic analysis of Phi-treated plants showed significant differences in the levels of many metabolites especially amino acids. Multivariate statistical approaches, such as principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), were employed to explore the relationships between metabolites to detect group differences. A good discrimination between the control and the Phi-treated plants was observed, which demonstrated that significant changes in the metabolite profile have been caused by the two different Phi applications (foliar or post-harvest). This finding suggests that the alteration of specific metabolite levels by accumulation of Phi can lead to resistance against the pathogen.
文摘An efficient and rapid Agrobacterium tumefaciens-mediated transformation protocol was developed to generate activation-tagged mutant lines with the aim of large-scale functional analysis of the potato genome. The explants were inoculated with an Agrobacterium strain harboring the binary plasmid pSKI074 containing four CaMV 35S enhancers in the T-DNA region which activates the downstream genes in the host plant after its integration. Various parameters investigated to increase transformation efficiency were the type and age of explant, cultivar, hormone combinations, preculture of explants, period of co-cultivation with bacteria and concentration of bacterial cultures used for transformation. Stem explants from 5 week old plantlets of cv. Bintje which had undergone phytohormone pretreatment for 4 days, inoculation with diluted bacterial concentration of OD600 = 0.2 containing acetosyringone followed by 2 days of co-cultivation and selection in media with IAA and trans-zeatin all helped in greatly improving the transformation efficiency. The total time required from infection to rooted shoots was 6-7 weeks. Initial evidence for stable integration and expression of the transgenes by PCR analysis showed that over 93% of the regenerated lines were transgenic and this was confirmed by Southern hybridization.
文摘Potato late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating diseases in the agricultural sector around the world. Many genes (R genes) conferring resistance to late blight have been identified in various potato species and most of these R genes have been used in potato breeding. The aim of this study was to develop and validate PCR-based assays for the R genes Rpi-blb1, Rpi-blb2, Rpi-blb3 and Rpi-bt1, to distinguish between late blight resistant and late blight susceptible potato progeny in the given breeding background. A total of 100 breeding progeny were screened for the presence of these R genes and tested for resistance against P. infestans mating type A2, genotype US-8 strain, using detached leaf and tuber rot assays. PCR products for the Rpi-blb1 and Rpi-bt1 resistance genes were identified in the resistant progeny but were absent in the susceptible ones;therefore these PCR assays could differentiate between late blight resistant and susceptible plants. Genotypic data from the DNA markers derived from the Rpi-blb1 and Rpi-bt1 genes was found to correlate with the phenotypic data for foliar late blight but not with data for tuber rot. Our results demonstrate that markers derived from these two R genes could be useful for marker-assisted selection (MAS) for foliar late blight resistance in potato breeding programs.
文摘Total phenolics, anthocyanins and antioxidant capacity of five coloured/pigmented (AR2009-10, Adirondack Red, Adirondack Blue, Congo, and POROIPG22-1) and two unpigmented potato genotypes (“Anuschka” and “Russet Burbank”) were assessed in fresh (with and without skin) and commercially processed/cooked/reconstituted products. Ascorbate profiles of the seven genotypes also were investigated using fresh tuber/tissue only. The results showed that genotypes greatly varied in their contents of bioactive compounds. Ascorbate profiles of the genotypes were not associated to any particular flesh colour/pigment. However, the pigmented potatoes had 1.5 to 2.5 times more the phenolics, 2 to 3 times more antioxidant capacity and higher levels of anthocyanins (13.98 to 38.57 mg C3GE-100g FW) compared to unpigmented genotypes. No anthocyanins were detected in the unpigmented potatoes. Significant losses of total phenolics, anthocyanins and total antioxidant capacity were found during peeling (18% - 23%), blanching process (40% - 60%) and further cooking/ reconstitution (7% - 12%) with no prominent genotype differences. Together, 65 to 90% of these bioactive compounds were lost during processing. The results suggested that pigmented potatoes contained higher amounts of total phenolics and anthocyanins and blanching step took away the most of the original bioactive compounds.
文摘In Brassicaceae, myrosinase catalyzes the hydrolysis of glucosinolate and plays an important role in anti-herbivore defense. We have cloned and characterized the full- length complementary DNA of myrosinase gene from Brassicaparachinensis that exhibits high sequence identity with myrosinase genes from other Brassica species. To investigate the role of this myrosinase in defense against the diamondback moth （Plutella xylostella）, we constructed an RNA-interference （RNAi） cassette expressing a double-stranded RNA that targeted myrosinase and transfected it into B. parachinensis. Myrosinase was sup- pressed in the resulting transgenic plants. Diamondback moth larvae feeding on transgenic plants had lower larval and pupal weights, longer pupal duration, and lower fecundity than those feeding on non-transgenic plants, suggesting that the diamondback moth has adapted to the glucosinolate-myrosinase defensive system. Therefore, the suppression of myrosinase is a potential approach for controlling the diamondback moth.
