Loquat (Eriobotrya japonica) is a model fruit for investigating flesh lignification during storage and response to chilling injury. However, the investigations of enzymes and coding genes and loquat fruit lignification under low-temperature storage are still limited. Here, the activity and transcript levels of up-stream enzymes of the phenylpropanoid pathway, including l-phenylalanine ammonia-lyase (PAL), cinnamate 4-hydroxylase (C4H), and 4-coumarate:coenzyme A ligase (4CL), were investigated. The results indicated that activity of these enzymes was positively correlated with loquat fruit lignification and suppression of these increases by heat treatment (HT) and low-temperature conditioning (LTC) significantly alleviated loquat fruit lignification. Coding genes for these enzymes were subsequently isolated based on information from an RNA-seq database and expression of Ej4CL1 was found to be the most responsive to low temperature and inhibition by HT and LTC treatment, whereas the other genes were less responsive to these treatments. Furthermore, function of Ej4CL1 was analyzed by transient overexpression in tobacco leaves, where it stimulated lignin accumulation. Ej4CL1 may be a key candidate that involved in CI-related loquat fruit lignification.
Xue Li, Chen Zang, Hang Ge, Jing Zhang, Donald Grierson, Xue-ren Yin, and Kun-song Chen
Bo Zhang, Xue-Ren Yin, Ji-Yuan Shen, Kun-Song Chen, and Ian B. Ferguson
The relationship between lipoxygenase (LOX) pathway-derived volatiles and LOX gene expression was evaluated in kiwifruit [Actinidia deliciosa (A. Chev.) C.F. Liang et A.R. Ferguson var. deliciosa cv. Bruno] during postharvest ripening at 20 °C. The C6 aldehydes n-hexanal and (E)-2-hexenal were abundant in peel compared with flesh tissue and declined as kiwifruit ripened. Esters such as ethyl butanoate and methyl butanoate were lower in the peel than flesh and accumulated when the fruit underwent a climacteric rise in ethylene production. Total LOX activity was higher in the peel than in the flesh and increased as kiwifruit ripened. Expression of AdLox2, AdLox3, AdLox4 and AdLox6 was high in the peel, whereas AdLox1 and AdLox5 showed similar levels in the peel and flesh at the ethylene climacteric. AdLox1 and AdLox5 transcript levels increased and AdLox2, AdLox3, AdLox4 and AdLox6 levels decreased during postharvest fruit ripening. Principal component analysis showed that n-hexanal and (E)-2-hexenal were grouped with LOX genes that were downregulated as kiwifruit ripened. The possible roles of LOX genes in relation to kiwifruit volatile formation during fruit ripening are discussed.
Song-jun Zeng, Zhi-lin Chen, Kun-lin Wu, Jian-xia Zhang, Cheng-ke Bai, Jaime A. Teixeira da Silva, and Jun Duan
Nothodoritis zhejiangensis Z. H. Tsi is a rare and endangered epiphytic orchid restricted to a narrow distribution in China. The species is threatened with extinction as a result of loss of suitable habitats. An efficient propagation system is part of this species’ conservation plan. Seed germination reached 64.7% on Knudson's C (KC) medium containing 1.0 mg·L−1 α-naphthaleneacetic acid (NAA), 10% coconut water, and 0.1% activated charcoal. After 50 days culture, most callus (71.3%) from seed-derived protocorms formed on KC medium with 1.0 mg·L−1 2,4-dichlorophenoxyacetic acid and 10% coconut water. Callus could be subcultured 12 times during ≈2 years with more than a 3.0-fold increase from the third to the twelfth subculture. Furthermore, 84% of callus from the tenth sub-culture on KC media supplemented with 1.0 mg·L−1 NAA, 5% coconut water, and 0.1% activated charcoal formed protocorm-like bodies (PLBs). Many (57%) protocorms on KC medium containing 1.0 mg·L−1 6-benzylaminopurine and 10% coconut water formed PLBs. Both callus and PLBs formed simultaneously from different protocorms on KC medium containing 0.01 to 1.0 mg·L−1 thidiazuron and 10% coconut water. PLBs were incubated 12 times during ≈2 years with an approximate 2.4-fold increase per sub-culture. Both callus and PLBs maintained their competence to regenerate plantlets. Hyponex N026 medium supplemented with 1.0 mg·L−1 NAA, 50 g·L−1 banana homogenate, and 0.1% activated charcoal was suitable for plantlet formation and growth of 95.5% of plantlets that developed from PLBs. The roots of plantlets 2 cm in height or taller were wrapped in Chilean sphagnum moss and fixed to a fir bark block; 69.3% of plantlets survived after 180 days in a greenhouse. This protocol is an efficient means for the large-scale propagation of this endangered orchid.