A method for promoting callus induced by stem segments of macadamia nuts

Abstract

The invention belongs to the technical field of macadamia ternifolia F.Muell. cultivation, in particular to a method for promoting callus induction of macadamia ternifolia F.Muell. stems. The method comprises the five steps of explant collection, explant pretreatment, disinfection, inoculation and culture. By means of a treatment scheme of the invention, the explant contamination rate is 5% or below; the callus induction rate is 96% or above; calluses and regenerated buds can be obtained in a short period of time; the calluses can be used as inducing materials for sterile buds; no disinfectiontreatment is needed; the regenerated buds grow faster; and a situation of cell tissue necrosis is not prone to occur.

Description

Technical field [0001] The invention belongs to the technical field of macadamia nut cultivation, in particular to a method for promoting the induction of callus from the stems of macadamia nuts. Background technique [0002] Macadamia (Macadamia ternifolia F.Muell.) belongs to the Proteaceae Juss. (Proteaceae Juss.) Macadamia (Macadamia F.Muell.) perennial evergreen tree fruit tree, also known as macadamia, Queensland chestnut, macadamia walnut, etc., native to Australia is rich in nutrients and has high economic and medicinal value. It enjoys the reputation of "the king of dried fruits" among many nuts; China first introduced it to the Taipei Botanical Garden for planting in 1910, and it was not until 1979. Carry out research on introduction, trial planting and cultivation technology; in recent years, China has introduced varieties from Australia to promote planting in Yunnan and Guangxi; with the improvement of people’s quality of life, macadamia has been developed in hot area...

AI Technical Summary

Problems solved by technology

[0003] Macadamia nuts are generally planted by asexual budding seedlings, and the seedlings are used as rootstocks, which take a long time to cultivate, and there is currently a lack of good rootstocks for economical and efficient rapid propagation in planting; in nurseries and orchards, grafting, cuttings, etc. are mainly used for propagation. However, the root system of seedlings propagated by cutting, grafting, etc. is underdeveloped, the plant grows slowly, and the rooting time is long. At the same time, cutting propagation requires pruning, shaping and other auxiliary management, which greatly limits the planting and industry of macadamia nuts in China. In addition, when macadamia nut stems are used as explants to induce callus or sterile buds, the explants usually collected have complex and diverse endophytes, which are difficult to disinfect, have a high pollution rate, and are easily necrotic. Carry out large-scale production; In recent years, the Subtropical Crops Research Institute of Guizhou Academy of Agricultural Sciences has introduced some macadamia nut varieties (788, H2, OC, etc.), which have great development prospects and economic benefits; but these varieties still lack an economic , Efficient rapid propagation system: The isolated stem segment has the advantages of short culture period, fast reproduction, detoxification, and no pollution, which can effectively reduce the risk of plant disease transmission and maintain the excellent traits of the mother plant. It is the most reliable way to achieve rapid propagation of macadamia nuts. An effective method; in vitro rapid propagation has been reported on other plants of the family Solanaceae such as longan, silver-leafed tree, and silver birch; at present, there are some isolated Reports on the rapid propagation of body tissues, such as the macadamia nut tissue culture propagation method disclosed in the application number 201710038704.1, the invention selects the young stems of macadamia nuts in the same year and inoculates them in the starting medium after disinfection treatment: 1 / 2MS+6-BA+0.5 ~2.0mg / L+IBA0.5~1.0mg / L+sucrose 30g / L+agar 4.6g / L; proliferation medium: 1 / 2MS+GA0.5~1.0mg / L+sucrose 30g / L+agar 4.6g / L; Rooting medium: 1 / 4MS+IBA1~5mg / L+sucrose 5~10g / L+agar 2g / L+0.5%PEG for rooting culture; another example is the method for rapid seedling growth of macadamia nut tissue culture disclosed in application number CN201710288673.5 In this invention, the clipped macadamia nut branches are soaked in mercuric chloride, washed with sterile water, and then inoculated with a medium: the first solid medium is 1 / 2MS, 7g / L agar, 15g / L sucrose, 2g / L L carbon powder, 15g / L molasses fermentation broth, 15mg / L peptone, pH 5.8; the second solid medium is 1 / 2MS, 2mg / L 6-benzylaminoadenine, 1.0mg / L red Mycin, 15g / L of agar, 15g / L of sucrose, 15g / L of molasses fermentation broth, 2g / L of inositol, 1mg / L of niacin, 80g / L of paclobutrazol, pH 5.8; the third solid The medium is 1 / 2MS, 2mg / L 6-benzylaminoadenine, 1.0mg / L gibberellin, 2mg / L indole butyric acid, 15g / L agar, 15g / L sucrose, 15g / L L of molasses fermentation broth, 1mg / L Thiamine hydrochloride, 2mg / L pyridoxine hydrochloride, 2g / L inositol, 1mg / L sodium molybdate, 5mg / L boric acid, 80mg / L disodium edetate, pH is 5.8; In addition, Guo Lingfei in the document "Preliminary Report on Tissue Culture Research of Macadamia Nuts" disclosed that macadamia nuts with bud stems were used as explants, washed with soapy water, then washed with running water, soaked in 75% ethanol, and then used Mercury chloride treatment, rinse with sterile water for several times; or soak quickly with mercury chloride at the sampling site, transfer to sterile water, then treat with mercury chloride, and finally rinse with sterile water several times; the initial culture medium is 1 / For 2MS medium, add 7g of agar and 30g of sucrose to each liter of medium, adjust the pH value to 5.8, and sterilize at 121°C and 107.87kPa; the subculture medium is 1 / 2MS medium, with different concentrations of BA and GA added ; These methods all have defects such as incomplete disinfection, high contamination rate, less callus obtained, weak differentiation ability, etc.