Optimization of protoplast preparation and regeneration conditions of Stemphylium lycopersici

doi:10.3969/j.issn.1004-1524.2022.02.15

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (2): 337-342.DOI: 10.3969/j.issn.1004-1524.2022.02.15

• Plant Protection • Previous Articles Next Articles

Optimization of protoplast preparation and regeneration conditions of Stemphylium lycopersici

GONG Weijie(), LIU Hong, HUANG Ping, WANG Hui, ZHOU Qian^*()

Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insects Pests, College of Plant Protection, Hunan Agricultural University, Changsha 410128, China

Received:2021-05-24 Online:2022-02-25 Published:2022-03-02
Contact: ZHOU Qian

Abstract

Abstract:

Protoplastpreparation conditions ofStemphylium lycopersiciwas optimizedusing strain CS 12 fromhypha age, enzyme species and concentration, enzyme combination, enzymolysis temperature and time, isotonic solutions, and the optimal regeneration medium. Theprotoplasm preparation and regeneration of S.lycopersici was successfully established by using mycelium cultured 36 h, 0.7 mol·L^-1 NaCl solution as the osmotic stabilizer, 10 mg·mL^-1Kitalase as enzymatic hydrolysate at 28℃ for 3.5 h, and the regeneration efficiency was the highest on the YEPS regeneration medium. The establishment of the protoplast preparation and regeneration of S.lycopersici laid foundation for further research on functional genomics and pathogenic mechanism of S.lycopersici.

Key words: Stemphylium lycopersici, protoplast, regeneration, osmotic stabilizer, hypha age

CLC Number:

S432.4⁺4

GONG Weijie, LIU Hong, HUANG Ping, WANG Hui, ZHOU Qian. Optimization of protoplast preparation and regeneration conditions of Stemphylium lycopersici[J]. Acta Agriculturae Zhejiangensis, 2022, 34(2): 337-342.

Figures/Tables 8

Fig.1 Effect of osmotic stabilizers on preparation of protoplasts

Fig.2 Effects of different enzyme types and concentrations on protoplasts preparation

Fig.3 Effect of different enzyme combinations on protoplasts preparation Kit, Kitalase; Lys, Lysing enzymes; Dri, Driselase; Sna, Snailase.

Fig.4 Effect of growth time of fungi on preparation of protoplasts

Fig.5 Effect of enzymolysis temperature on preparation of protoplasts

Fig.6 Effect of enzymolysis time on preparation of protoplasts

Fig.7 Protoplast of Stemphylium lycopersici CS12

Fig.8 Regeneration rate of protoplast in different medium

References 18

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Optimization of protoplast preparation and regeneration conditions of Stemphylium lycopersici

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