Ric-Varas Pablo, Barceló Marta, Rivera Juan A, Cerezo Sergio, Matas Antonio J, Schückel Julia, Knox J Paul, Posé Sara, Pliego-Alfaro Fernando, Mercado José A
Instituto de Hortofruticultura Subtropical y Mediterránea "La Mayora" (IHSM-UMA-CSIC), Departamento de Botánica y Fisiología Vegetal, Universidad de Málaga, 29071 Málaga, Spain.
IFAPA Centro de Málaga, Cortijo de la Cruz s/n, 29140 Málaga, Spain.
Plants (Basel). 2020 Jun 27;9(7):805. doi: 10.3390/plants9070805.
Cell cultures derived from strawberry fruit at different developmental stages have been obtained to evaluate their potential use to study different aspects of strawberry ripening. Callus from leaf and cortical tissue of unripe-green, white, and mature-red strawberry fruits were induced in a medium supplemented with 11.3 µM 2,4-dichlorophenoxyacetic acid (2,4-D) under darkness. The transfer of the established callus from darkness to light induced the production of anthocyanin. The replacement of 2,4-D by abscisic acid (ABA) noticeably increased anthocyanin accumulation in green-fruit callus. Cell walls were isolated from the different fruit cell lines and from fruit receptacles at equivalent developmental stages and sequentially fractionated to obtain fractions enriched in soluble pectins, ester bound pectins, xyloglucans (XG), and matrix glycans tightly associated with cellulose microfibrils. These fractions were analyzed by cell wall carbohydrate microarrays. In fruit receptacle samples, pectins were abundant in all fractions, including those enriched in matrix glycans. The amount of pectin increased from green to white stage, and later these carbohydrates were solubilized in red fruit. Apparently, XG content was similar in white and red fruit, but the proportion of galactosylated XG increased in red fruit. Cell wall fractions from callus cultures were enriched in extensin and displayed a minor amount of pectins. Stronger signals of extensin Abs were detected in sodium carbonate fraction, suggesting that these proteins could be linked to pectins. Overall, the results obtained suggest that fruit cell lines could be used to analyze hormonal regulation of color development in strawberry but that the cell wall remodeling process associated with fruit softening might be masked by the high presence of extensin in callus cultures.
已获得来自不同发育阶段草莓果实的细胞培养物,以评估其在研究草莓成熟不同方面的潜在用途。在添加了11.3 µM 2,4-二氯苯氧乙酸(2,4-D)的培养基中,于黑暗条件下诱导未成熟绿色、白色和成熟红色草莓果实的叶片和皮层组织产生愈伤组织。将已建立的愈伤组织从黑暗转移到光照条件下可诱导花青素的产生。用脱落酸(ABA)替代2,4-D可显著增加绿色果实愈伤组织中的花青素积累。从不同果实细胞系以及处于同等发育阶段的果实花托中分离细胞壁,并依次分级分离以获得富含可溶性果胶、酯结合果胶、木葡聚糖(XG)以及与纤维素微纤丝紧密相关的基质聚糖的级分。这些级分通过细胞壁碳水化合物微阵列进行分析。在果实花托样品中,所有级分中果胶都很丰富,包括那些富含基质聚糖的级分。果胶含量从绿色阶段到白色阶段增加,随后这些碳水化合物在红色果实中溶解。显然,白色和红色果实中XG含量相似,但红色果实中半乳糖基化XG的比例增加。愈伤组织培养物的细胞壁级分富含伸展蛋白,且显示出少量果胶。在碳酸钠级分中检测到更强的伸展蛋白抗体信号,表明这些蛋白质可能与果胶相连。总体而言,所得结果表明果实细胞系可用于分析草莓颜色发育的激素调控,但与果实软化相关的细胞壁重塑过程可能会被愈伤组织培养物中高含量的伸展蛋白所掩盖。