[ Objective ] The paper was to explore effects of Tomato aspermy virus （TAV） -free on chlorophyll content and nitrogen metabolism of Chrysanthemum morifolium Ramat. ＇Huaibai＇, in order to provide a theoretical bas...[ Objective ] The paper was to explore effects of Tomato aspermy virus （TAV） -free on chlorophyll content and nitrogen metabolism of Chrysanthemum morifolium Ramat. ＇Huaibai＇, in order to provide a theoretical basis for field popularization and application of TAV-free ＇ Huai bai＇ plantlets. [ Method] With previously acquired TAV-free test-tube plantlets of ＇ Huaibai＇ as the experimental materials and its ordinary test-tube plantlets as the control （CK）, chlorophyll content and some indexes related to nitrogen metabolism in leaves were determined during subculture. [ Result] During subculture, the contents of chlorophyll a and chlorophyll b in leaves of CK and TAV-free ＇ Huaibai＇ test-tube plantlets increased with the subculture days. Both realized the largest increase at the 22＇h day of subculture, and their increase amplitudes were 5.24% and 41.69%, respectively. Compared with CK, the chlorophyll a and chlorophyll b contents of TAV-free test-tube plantlets increased at the same time point of subculture, with significant difference （P 〈0.05） or extremely significant difference （P 〈0.01 ）. Further- more, the nitrate reductase （NR） activity, free proline and soluble protein contents of TAV-free ＇ Huaibai＇ were all significantly higher than those of CK, and the highest increase amplitudes were 27.99% （ subculture for 22 d）, 28.10% （ subculture for 22 d） and 5.89% （ subculture for 38 d）, respectively. [ Conclusion] TAV-free could effectively improve chlorophyll content and nitrogen metabolism level of ＇ Huaibai＇ , and was conducive to its growth and development.展开更多
Poor resolution for the identification of endopeptidase(EP) activity in activity gel assays is a critical issue in the analysis of the postharvest physiology of rose petals. In this paper, major factors influencing EP...Poor resolution for the identification of endopeptidase(EP) activity in activity gel assays is a critical issue in the analysis of the postharvest physiology of rose petals. In this paper, major factors influencing EP activity gel assays were evaluated. The results showed that a phosphate(NaH_2PO_4/Na_2HPO_4) buffer favors the detection of clear EP bands, as compared to Tris–HCl buffer. Removal of salts and pigments with Sephadex G-25 columns was vital to the measurement of EP activity in rose petal extracts. For optimal resolution of bands, we show that before electrophoresis,samples should be treated for 10 min at 40 °C. Additionally, electrophoresis should be done in 12% SDS–PAGE co-polymerized with 0.15%(w/v) gelatin. After electrophoresis, the optimal incubation temperature and pH are 42 °C and 7.0, respectively. Using our optimized assay, Rh-EP1,Rh-EP2, Rh-EP3, three proteases with molecular masses of 200, 123.5, and 97.4 kD, respectively, were detected in rose petals. Experiments using EP class-specific inhibitors revealed that Rh-EP2 and Rh-EP3 were both serine proteases, while Rh-EP1 was a metalloprotease. In this study, we also measured changes in EP activity during flower opening, senescence, and water deficit stress(WDS) using our optimized activity gel assay,and found that Rh-EP3 may be more relevant to senescence in roses compared with Rh-EP1 or Rh-EP2. Changes occurring to EPs after WDS were similar to those during the period from flower opening to senescence, and Rh-EP3 activities were greatly increased by WDS treatment. Collectively, our results suggest that significant increases in Rh-EPs activities, especially increases in Rh-EP3 activity, may contribute to the flower senescence induced under WDS treatment.展开更多
基金Supported by National Natural Science Foundation of China(31372105)Subproject of Special Research of National Traditional Chinese Medicine Industry(201407005-08)+1 种基金Support Project of Science and Technology Innovation Teams in Colleges and Universities of Henan Province(15IRTSTHN020)Project for Construction of Innovative Science and Technology Talent Team in Henan Province(C20130037)
文摘[ Objective ] The paper was to explore effects of Tomato aspermy virus （TAV） -free on chlorophyll content and nitrogen metabolism of Chrysanthemum morifolium Ramat. ＇Huaibai＇, in order to provide a theoretical basis for field popularization and application of TAV-free ＇ Huai bai＇ plantlets. [ Method] With previously acquired TAV-free test-tube plantlets of ＇ Huaibai＇ as the experimental materials and its ordinary test-tube plantlets as the control （CK）, chlorophyll content and some indexes related to nitrogen metabolism in leaves were determined during subculture. [ Result] During subculture, the contents of chlorophyll a and chlorophyll b in leaves of CK and TAV-free ＇ Huaibai＇ test-tube plantlets increased with the subculture days. Both realized the largest increase at the 22＇h day of subculture, and their increase amplitudes were 5.24% and 41.69%, respectively. Compared with CK, the chlorophyll a and chlorophyll b contents of TAV-free test-tube plantlets increased at the same time point of subculture, with significant difference （P 〈0.05） or extremely significant difference （P 〈0.01 ）. Further- more, the nitrate reductase （NR） activity, free proline and soluble protein contents of TAV-free ＇ Huaibai＇ were all significantly higher than those of CK, and the highest increase amplitudes were 27.99% （ subculture for 22 d）, 28.10% （ subculture for 22 d） and 5.89% （ subculture for 38 d）, respectively. [ Conclusion] TAV-free could effectively improve chlorophyll content and nitrogen metabolism level of ＇ Huaibai＇ , and was conducive to its growth and development.
基金supported by grants from the National Natural Science Foundation of China(30972025,31372105)
文摘Poor resolution for the identification of endopeptidase(EP) activity in activity gel assays is a critical issue in the analysis of the postharvest physiology of rose petals. In this paper, major factors influencing EP activity gel assays were evaluated. The results showed that a phosphate(NaH_2PO_4/Na_2HPO_4) buffer favors the detection of clear EP bands, as compared to Tris–HCl buffer. Removal of salts and pigments with Sephadex G-25 columns was vital to the measurement of EP activity in rose petal extracts. For optimal resolution of bands, we show that before electrophoresis,samples should be treated for 10 min at 40 °C. Additionally, electrophoresis should be done in 12% SDS–PAGE co-polymerized with 0.15%(w/v) gelatin. After electrophoresis, the optimal incubation temperature and pH are 42 °C and 7.0, respectively. Using our optimized assay, Rh-EP1,Rh-EP2, Rh-EP3, three proteases with molecular masses of 200, 123.5, and 97.4 kD, respectively, were detected in rose petals. Experiments using EP class-specific inhibitors revealed that Rh-EP2 and Rh-EP3 were both serine proteases, while Rh-EP1 was a metalloprotease. In this study, we also measured changes in EP activity during flower opening, senescence, and water deficit stress(WDS) using our optimized activity gel assay,and found that Rh-EP3 may be more relevant to senescence in roses compared with Rh-EP1 or Rh-EP2. Changes occurring to EPs after WDS were similar to those during the period from flower opening to senescence, and Rh-EP3 activities were greatly increased by WDS treatment. Collectively, our results suggest that significant increases in Rh-EPs activities, especially increases in Rh-EP3 activity, may contribute to the flower senescence induced under WDS treatment.
