Method for acquiring suspension cell line with hypocotyl of cabbage type rape as explant and application of suspension cell line

Abstract

The invention specifically relates to a method for acquiring a suspension cell line with the hypocotyl of cabbage type rape as explant and application of the suspension cell line, belonging to the technical field of plant cell culture. According to the invention, the aseptic hypocotyl of cabbage type rape is used as explant and subjected to induction of embryonic callus, culture of suspension cells and the like so as to construct the suspension cell line of the cabbage type rape; and the suspension cell line is used for research on and comparison of the different morphologies and activities ofsuspension cells under the conditions of normal supply of essential micronutrient elements and shortage of the essential micronutrient elements. The prepared suspension cells of the cabbage type rapeare high in activity, stable and uniform in morphology, short in culture period, not limited by natural environmental factors like season and climate, and applicable to research on the cell development of the cabbage type rape at a single cell level and the cell growth, morphology, transmembrane nutrient transport, physiological and biochemical metabolism and gene expression of the cabbage type rape under the condition of abiotic stress.

Description

technical field [0001] The invention belongs to the technical field of plant cell culture, and in particular relates to a method for obtaining a suspension cell line by taking the hypocotyl of Brassica napus as an explant and its application. Background technique [0002] Brassica napus L., Brassica napus, is one of the most important oil crops in the world. In my country, it has become the fifth largest crop after rice, corn, wheat, and soybean, and it is also the largest oil crop. , the annual planting area is about 6.7 million hm 2 , widely distributed in the Yangtze River Basin of my country, with a total output of more than 11 million tons, and an annual output of about 4.5 million tons of rapeseed oil, accounting for more than 40% of the total self-produced vegetable oil and more than 57% of the total herbal vegetable oil. Traditionally, the research on Brassica napus is based on plants. However, the complexity of the plants themselves leads to slow research progress. ...

AI Technical Summary

Problems solved by technology

Traditionally, the research on Brassica napus is based on plants. However, the complexity of the plants themselves leads to slow research progress. Suspension cells as single cells avoid the overall complexity of the plants.

In recent years, single-cell research has been widely used in other species, but Brassica napus still lacks an efficient and simple single-cell culture technology

[0003] Suspension cell has been widely concerned as a single cell, and the traditional Brassica napus suspension cell culture system is obtained by using microspore culture (Orr W., Keller W.A. and Singh J.1986, J.PlantPhysiol.126: 23-32), and then the suspension cell line obtained by the microspore culture method promoted the research of Brassica napus to a certain extent (Carswell, M.C., Grant, B.R. and Plaxton, W.C., 1997.Planta, 203(1), pp.67-74; Moraes, T.F. and Plaxton, W.C., 2000. European Journal of Biochemistry, 267(14), pp.4465-4476; Palma, D.A., Blumwald, E. and Plaxton, W.C., 2000. FEBS Letters, 486 (2), pp.155-158; Smith, C.R., Knowles, V.L. and Plaxton, W.C., 2000. European Journal of Biochemistry, 267(14), pp.4477-4485; Plaxton, W.C., Smith, C.R.and Knowles, V.L., 2002. Archives of biochemistry and biophysics, 400(1), pp.54-62; Singh, V.K., Wood, S.M., Knowles, V.L. and Plaxton, W.C., 2003. Planta, 218(2), pp.233- 239.), but the suspension cell line obtained by the method of microspore culture has the following disadvantages: (1) it is easily restricted by the season of material collection, Brassica napus blooms once a year, and cannot be sampled and cultivated at any time; Microscopic examination is to determine the suitable pollen development period, which is inconvenient to operate, and it is easy to miss the best period of pollen development; (3) The physiological state of the parent plant has a direct impact on the frequency of callus induction. In the season when the climate environment is unstable, There are significant differences in the callus induction rate of rapeseed microspores

8(1): 100~104; Yang Yuhua et al., 2002, Journal of Hubei Agricultural University. 22(4): 289-292; Yang Lu, 2012, Huazhong Agricultural University master's thesis; Hua, Y et al., 2016, Plant , cell&environment, 39(7), p.1601.), overcome the disadvantages of inconvenient microspore culture operation and being easily limited by seasons and environmental factors, but from the published articles, the cell line established by this method is dispersed Poor degree, severe deformity, and extremely low viability, it is difficult to provide excellent biological materials for single-cell study of Brassica napus

[0005] At present, there is no report on the successful use of Brassica napus hypocotyls as explants to obtain suspension cell lines. Therefore, it is necessary to establish a suspension cell line in Brassica napus that overcomes the above problems, called Brassica napus cells A level study offers new avenues

Images