关键词：
Brassica oleracea L. var. italica;Selenium;Biofortified;Bioactive substance;Sulforaphane;Glucosinolate;Myrosinase
摘要：
The effects of S (as sulphate) and Se (as selenite) treatment (S mM/Se mu M: 1/0, 1/50, 1/100, 1/150, 4/0, 4/50, 4/100, and 4/150) on the production of sulforaphane (an anticancer compound), the accumulation of its precursor substance, and the expression of genes related to glucoraphanin biosynthesis in broccoli were examined. Sulforaphane yield and myrosinase activity increased significantly with the combined application of 4 mM S and 100 mu M Se on broccoli. Furthermore, the concentrations of glucoraphanin (a sulforaphane precursor) and methionine (a glucoraphanin substrate) slightly changed after Se application. And the strong anticancer activity of compound Se-SMC was further improved. Analysis of related gene expression showed that MY, which encodes myrosinase, was strongly induced by Se treatment. Thus, the myrosinase activity induced by Se treatment is the dominant factor affecting sulforaphane yield from glucoraphanin hydrolyzation. Selenium-sulfur biofortification provides a technical support for the cultivation of broccoli with high sulforaphane and high anti-cancer selenium compounds.
摘要：
The effects of S and Se treatment on cabbage, especially the interactions of S and Se metabolism with the biosynthesis of glucosinolate (GSL), including glucoraphanin, which is a major aliphatic GSL in cruciferous vegetables and the precursor of the anticancer compound sulforaphane, were examined. Cabbage plants were treated with sulfate and selenite (SeO32-), and the total S, Se, and GSL contents of cabbage head and outer foliage leaves were measured. Results showed that selenite treatment was beneficial to GSL biosynthesis and Se accumulation in cabbage head and outer foliage leaves. GSL synthesis was induced by exogenous selenite-elevated sulfate treatment at certain concentration ratios, i.e., 50-mu M selenite + 1-mM sulfate or 100-mu M selenite + 4-mM sulfate. A high exogenous sulfate concentration was more favorable to GSL accumulation than a low sulfate concentration. According to the relative expression of genes on GSL synthesis, an increase in the GSL content was attributed to the upregulation of gene expression and possible transportation from the outer foliage leaf to the head of cabbage. These results might be helpful for increasing the health benefits of cabbage by supplying exogenous S and Se. Further research should explore the effects of sulfate and selenite on GSL precursor substances to reveal the reason why total GSL contents increased.
摘要：
In this study, a new SYBR Green qPCR (qRT-PCR) and a nested RT-PCR (nRT-PCR) were developed to detect Potato mop-top virus (PMTV) in potato tuber tissues. The SYBR Green qRT-PCR and nRT-PCR assays were approximately 10(4)- and 10(3)-fold more sensitive than the conventional RT-PCR assay. The progeny tubers derived from PMTV-infected potato tubers were tested by conventional RT-PCR, SYBR Green qRT-PCR and nRT-PCR assays. Of the 17 samples, 9 (52.9%) were positive for PMTV by conventional RT-PCR, 11 (64.7%) were positive by nRT-PCR, and 17 (100%) were positive by SYBR Green qRT-PCR. Compared to nRT-PCR, SYBR Green qRT-PCR was showed to be more sensitive. The progeny plants exhibited foliar symptoms including chlorosis and reduction in leaf size when the PMTV-positive tubers were planted in a growth chamber at 20-22 degrees C. These findings suggest that PMTV has been passed on to the progeny plants and tubers.
关键词：
In vitro;Potato;PVA;Thermotherapy;Virus eradications
摘要：
With the aim of developing an effective protocol for virus elimination from potato (Solanum tuberosum L.) plantlets, thermotherapy coupled with isolating the first nodal cuttings by in vitro culture was successful to potato virus A (PVA) elimination. The survival ratio of potato plantlets was affective by thermotherapy temperatures and durations. The optimal thermotherapy temperature was 36±1 oC with highest survival ratio and effective elimination. The results of RT-PCR indicated that the regenerated plantlets obtained from the first cycle (four weeks) of thermotherapy in daytime at 36±1 oC with light intensity 40 μmole/m/s for 12 hr, and 20±1 oC in darkness for 12 hr had PVA infected. While isolated the first nodal cuttings and followed by thermotherapy at the first cycle conditions for another two weeks, the PVA could be eliminated. Thermotherapy was given by culturing the nodal cutting from the infected of PVA for six weeks in total on MS medium, and the PVA-free plantlets were obtained. In concluded that the protocol of thermotherapy coupled with isolating the first nodal cuttings by in vitro culture in the study can be effectively used for virus free plantlets in potato, and probably also for other vegetable propagated plant species.
摘要：
Broccoli (Brassica oleracea L. var. italica planck) has been highly valued because of its nutrient content, which has been attributed to both sulforaphane, a sulfur-containing compound, and selenium enrichment ability. The sulforaphane synthesis may be affected by Se/S metabolism because sulfate and selenate share the same initial pathway for uptake in plants. Selenocysteine methyltransferase (SMT) plays a critical role in the Se/S metabolism system. In the present work, we analyzed the SMT expression level and sulforaphane content in transgenic broccoli under the conditions of overexpression and RNAi knockdown of SMT under normal and selenium-stressed conditions. The relative expression value of SMT in the overexpression line is 13.93 % higher than that in the untransformed control. Interestingly, the sulforaphane content of the overexpression line was 14.09 % lower after selenium treatment, while that of the empty vector (pCAMBIA1301) transformed and untransformed plants was 59.04 and 66.56 % lower than that of the non-selenium-treated, respectively. In the SMT-RNAi line, which has a relative expression value for SMT 15.60 % lower than that of the untransformed, we cannot detect sulforaphane after selenium treatment. These results showed that SMT plays a key role in sulforaphane synthesis in a selenium-rich environment. Specifically, overexpression of SMT decreases the negative effect of selenium on sulforaphane synthesis, while knockdown of SMT by RNAi enhances the negative effect.
