Microorganism DNA of rhizosphere soil from Pinus koraiensis and Pinus sylvestriformis were extracted by proteinase K based on SDS method, CTAB method, PVP (polyvinylpolypyrrolidone) method, and freezing and thawing ...Microorganism DNA of rhizosphere soil from Pinus koraiensis and Pinus sylvestriformis were extracted by proteinase K based on SDS method, CTAB method, PVP (polyvinylpolypyrrolidone) method, and freezing and thawing method and the crude DNA from rhizosphere soil were purified by dialysis method, silver beads absorption method, and squeezing DNA gel method. The results of different extracting and purifying methods were compared and evaluated. Results indicated that the best method of extraction for microorganism DNA in rhizosphere soil was proteinse K based on SDS method with high salt concentration of 1.0% (w/v) NaCl, which could effectively eliminate humic acids and other impurities. The dialysis method was suitable to purify DNA from rhizosphere soil because of effectively removing brown matters and humic acids and the purified products were suited to PCR amplification. Squeezing DNA gel method was also a good purification method with the advantage of inexpensive in cost and efficient in use.展开更多
A glasshouse experiment using a rhizobox technique was conducted to examine salt dynamics in the rhizosphere of a salt-tolerant grass, Puccinellia ciliata Bor. ’Irwin Hunter’, grown in a loamy soil, and to study the...A glasshouse experiment using a rhizobox technique was conducted to examine salt dynamics in the rhizosphere of a salt-tolerant grass, Puccinellia ciliata Bor. ’Irwin Hunter’, grown in a loamy soil, and to study the effect of rainfall flush on salt accumulation in the rhizosphere. The rhizobox (10 × 5.5 × 50 cm) had a nylon mesh (1 μm) positioned vertically in the middle to create two compartments filled with soil amended with 1 g NaCl kg-1. The plants were grown in one compartment only. Flushed treatments received 275 mL of deionized water two days before harvest. In the plant-growing compartment, soils were sectioned vertically at 5 cm intervals. Significant differences in soil electrical conductivity (EC) (P < 0.05) and pH (P < 0.05) were observed for depths, but not between flushed and non-flushed treatments. In the no-plant compartment (rhizosphere), soil cores were taken horizontally at depths of 5, 20 and 40 cm and sliced at 1, 2, 3, 4, 5, 7, 10, 15 and 20 mm away from the roots. Soil EC and Cl- concentration at the 5 and 20 cm depths, and Na+ concentration at the 5 cm depth significantly decreased (P < 0.05) with the distance away from the root, but no significant differences were observed in soil pH and concentrations of the K+ and Ca2+. The flush treatment only had significant influence on soil EC, pH, and Cl- concentration at the 20 cm depth. Thus, salt accumulation could occur in the rhizosphere of salt-tolerant species on saline soils, and the periodic low rainfall might not have a strong influence on salt distribution in the rhizosphere and/or root zone.展开更多
基金This project was supported by the Knowledge Innovation Project from Chinese Academy (KZCXI-SW-01) and the National Natural Science Foundation of China (30070158).
文摘Microorganism DNA of rhizosphere soil from Pinus koraiensis and Pinus sylvestriformis were extracted by proteinase K based on SDS method, CTAB method, PVP (polyvinylpolypyrrolidone) method, and freezing and thawing method and the crude DNA from rhizosphere soil were purified by dialysis method, silver beads absorption method, and squeezing DNA gel method. The results of different extracting and purifying methods were compared and evaluated. Results indicated that the best method of extraction for microorganism DNA in rhizosphere soil was proteinse K based on SDS method with high salt concentration of 1.0% (w/v) NaCl, which could effectively eliminate humic acids and other impurities. The dialysis method was suitable to purify DNA from rhizosphere soil because of effectively removing brown matters and humic acids and the purified products were suited to PCR amplification. Squeezing DNA gel method was also a good purification method with the advantage of inexpensive in cost and efficient in use.
文摘A glasshouse experiment using a rhizobox technique was conducted to examine salt dynamics in the rhizosphere of a salt-tolerant grass, Puccinellia ciliata Bor. ’Irwin Hunter’, grown in a loamy soil, and to study the effect of rainfall flush on salt accumulation in the rhizosphere. The rhizobox (10 × 5.5 × 50 cm) had a nylon mesh (1 μm) positioned vertically in the middle to create two compartments filled with soil amended with 1 g NaCl kg-1. The plants were grown in one compartment only. Flushed treatments received 275 mL of deionized water two days before harvest. In the plant-growing compartment, soils were sectioned vertically at 5 cm intervals. Significant differences in soil electrical conductivity (EC) (P < 0.05) and pH (P < 0.05) were observed for depths, but not between flushed and non-flushed treatments. In the no-plant compartment (rhizosphere), soil cores were taken horizontally at depths of 5, 20 and 40 cm and sliced at 1, 2, 3, 4, 5, 7, 10, 15 and 20 mm away from the roots. Soil EC and Cl- concentration at the 5 and 20 cm depths, and Na+ concentration at the 5 cm depth significantly decreased (P < 0.05) with the distance away from the root, but no significant differences were observed in soil pH and concentrations of the K+ and Ca2+. The flush treatment only had significant influence on soil EC, pH, and Cl- concentration at the 20 cm depth. Thus, salt accumulation could occur in the rhizosphere of salt-tolerant species on saline soils, and the periodic low rainfall might not have a strong influence on salt distribution in the rhizosphere and/or root zone.