Patent
An Exiguobacterium indicum YAN2 having high-concentration selenite tolerance, the preservation number thereof being GDMCC No. 61594, and a method for fermenting inorganic selenium into nano-selenium by using the strain are provided. The obtained nano-selenium can promote the growth of leaf vegetables, increase the content of chlorophyll, Vc, and total phenol, etc, reduce the content of nitrate, and improve the quality of leaf vegetables. In addition, the present application further provides a nano-selenium liquid fertilizer prepared by fermentation using the strain.
1. A method for synthesizing nano-selenium, comprising:
1) inoculating Exiguobacterium indicum with preservation number GDMCC NO. 61594 into a lysogeny broth (LB) liquid medium, and performing activated culture until OD 600 is approximately equal to 1.0 to obtain activated bacteria liquid;
2) inoculating the activated bacteria liquid obtained in step 1) into a fermentation tank containing a fermentation medium at an amount of inoculation by a volume ratio of 2.5±0.2%, and carrying out fermentation until the bacterial cells content is 0.4-0.6 mg/mL to obtain fermented bacteria liquid,
wherein the fermentation medium is prepared by: adding up to 1 L deionized water to 5 g of yeast extract, 10 g of tryptone, and 10 g of NaCl; sterilizing, and then adding 1 M selenite solution to make a final concentration of 5 mM selenite in the medium; and
3) centrifuging the fermented bacteria liquid obtained in step 2), and collecting a precipitate that is the nano-selenium.
2. The method according to claim 1 , wherein the fermentation in step 2) refers to a fermentation temperature of 30±1° C., a stirring speed of 150±5 rpm, and a tank pressure being 1.0±0.4 F/cm 2 .
3. A method for preparing a nano-selenium liquid fertilizer, comprising:
1) inoculating Exiguobacterium indicum with preservation number GDMCC NO. 61594 into an LB liquid medium, and performing activated culture until OD 600 is approximately equal to 1.0 to obtain activated bacteria liquid;
2) inoculating the activated bacteria liquid into a fermentation tank containing a selenium-containing terrific broth (TB) fermentation medium at an amount of inoculation of 2.5±0.2%, and carrying out fermentation to obtain a fermented liquid fertilizer,
wherein the selenium-containing TB fermentation medium is prepared by adding up to 1 L deionized water to 24 g/L of a yeast extract, 20 g/L of tryptone, 4 ml/L of glycerol, 72 mM K 2 HPO 4 , 17 mM KH 2 PO 4 , and 5 mM Na 2 SeO 3 ; and
3) centrifuging the fermented liquid fertilizer, and collecting precipitate, washing with sterile normal saline for 3-4 times, and then resuspending the precipitate with sterile water in 1/20 of a volume of the fermented liquid fertilizer, to obtain the nano-selenium liquid fertilizer.
4. The method according to claim 3 , wherein the fermentation in step 2) refers to a fermentation temperature of 30±1° C., a stirring speed of 150±5 rpm, and a tank pressure being 1.2±0.2 F/cm 2 .