Hannachi Sami, Werbrouck Stefaan, Bahrini Insaf, Abdelgadir Abdelmuhsin, Siddiqui Hira Affan, Van Labeke Marie Christine
Department of Biology, College of Science, University of Hail, P.O. Box 2440, Hail 81451, Saudi Arabia.
Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000 Ghent, Belgium.
Plants (Basel). 2021 Nov 22;10(11):2539. doi: 10.3390/plants10112539.
An efficient regeneration protocol was applied to regenerate shoots on salt stress-tolerant calli lines of aubergine (). These NaCl-tolerant cell lines were obtained by two different methods. On the one hand, the developed callus tissue was transferred to a medium with a continuous salt content of 40, 80, 120, or 160 mM NaCl. On the other hand, the callus tissue was subjected to a stepwise increasing salinity to 160 mM NaCl every 30 days. With the second method, calli which could be selected were characterized by compact growth, a greenish color, and absence of necrotic zones. When grown on salt-free medium again, NaCl-tolerant calli showed a decline in relative growth rate and water content in comparison to the control line. This was more obvious in the 120 mM NaCl-tolerant callus. Lipid peroxidase activity increased in 40 and 80 mM NaCl-tolerant calli; yet did not increase further in 120 mM-tolerant callus. An increase in ascorbic acid content was observed in 80 and 120 mM NaCl-tolerant calli compared to the 40 mM NaCl-tolerant lines, in which ascorbic acid content was twice that of the control. All NaCl-tolerant lines showed significantly higher superoxide dismutase (SOD) (208-305-370 µmol min mg FW) and catalase (CAT) (136-211-238 µmol min mg FW) activities compared to control plants (231 and 126 µmol min mg FW). Plants were regenerated on the calli lines that could tolerate up to 120 mM NaCl. From the 32 plants tested in vitro, ten plants with a higher number of leaves and root length could be selected for further evaluation in the field. Their high salt tolerance was evident by their more elevated fresh and dry weight, their more increased relative water content, and a higher number and weight of fruits compared to the wild-type parental control. The presented work shows that somaclonal variation can be efficiently used to develop salt-tolerant mutants.
一种高效的再生方案被应用于茄子耐盐愈伤组织系上再生芽。这些耐氯化钠的细胞系通过两种不同方法获得。一方面，将发育的愈伤组织转移到氯化钠含量持续为40、80、120或160 mM的培养基上。另一方面，愈伤组织每30天逐步增加盐度至160 mM氯化钠。采用第二种方法时，可选择的愈伤组织具有紧密生长、呈绿色且无坏死区的特征。当再次在无盐培养基上生长时，耐氯化钠愈伤组织与对照系相比，相对生长速率和含水量下降。这在耐120 mM氯化钠的愈伤组织中更为明显。脂质过氧化物酶活性在耐40和80 mM氯化钠的愈伤组织中增加；但在耐120 mM的愈伤组织中未进一步增加。与耐40 mM氯化钠系相比，耐80和120 mM氯化钠的愈伤组织中抗坏血酸含量增加，其中耐40 mM氯化钠系的抗坏血酸含量是对照的两倍。所有耐氯化钠系与对照植株（分别为231和126 μmol min mg FW）相比，超氧化物歧化酶（SOD）（208 - 305 - 370 μmol min mg FW）和过氧化氢酶（CAT）（136 - 211 - 238 μmol min mg FW）活性显著更高。在能耐受高达120 mM氯化钠的愈伤组织系上再生植株。在体外测试的32株植株中，可选择10株叶片数量较多且根长较长的植株用于田间进一步评估。与野生型亲本对照相比，它们较高的耐盐性表现为鲜重和干重更高、相对含水量增加更多以及果实数量和重量更高。所呈现的工作表明体细胞克隆变异可有效地用于培育耐盐突变体。
