Genome-wide identification of the CONSTANS-LIKE(COL)family and mechanism of fruit senescence regulation by PpCOL8 in sand pear(Pyrus pyrifolia)

Pyrus pyrifolia Nakai‘Whangkeumbae'is a sand pear fruit with excellent nutritional quality and taste.However,the industrial development of pear fruit is significantly limited by its short shelf life.Salicylic acid(SA),a well-known phytohormone,can delay fruit senescence and improve shelf life.However,the mechanism by which SA regulates CONSTANS-LIKE genes(COLs)during fruit senescence and the role of COL genes in mediating fruit senescence in sand pear are poorly understood.In this study,22 COL genes were identified in sand pear,including four COLs(Pp COL8,Pp COL9a,Pp COL9b,and Pp COL14)identified via transcriptome analysis and 18 COLs through genome-wide analysis.These COL genes were divided into three subgroups according to the structural domains of the COL protein.Pp COL8,with two B-box motifs and one CCT domain,belonged to the first subgroup.In contrast,the other three Pp COLs,Pp COL9a,Pp COL9b,and Pp COL14,with similar conserved protein domains and gene structures,were assigned to the third subgroup.The four COLs showed different expression patterns in pear tissues and were preferentially expressed at the early stage of fruit development.Moreover,the expression of Pp COL8 was inhibited by exogenous SA treatment,while SA up-regulated the expression of Pp COL9a and Pp COL9b.Interestingly,Pp COL8 interacts with Pp MADS,a MADS-box protein preferentially expressed in fruit,and SA up-regulated its expression.While the production of ethylene and the content of malondialdehyde(MDA)were increased in Pp COL8-overexpression sand pear fruit,the antioxidant enzyme(POD and SOD)activity and the expression of Pp POD1 and Pp SOD1 in the sand pear fruits were down-regulated,which showed that Pp COL8 promoted sand pear fruit senescence.In contrast,the corresponding changes were the opposite in Pp MADS-overexpression sand pear fruits,suggesting that Pp MADS delayed sand pear fruit senescence.The co-transformation of Pp COL8 and Pp MADS also delayed sand pear fruit senescence.The results of this study revealed that Pp COL8 can play a key role in pear fruit senescence by interacting with Pp MADS through the SA signaling pathway.

Involvement of long non-coding RNAs in pear fruit senescence under high-and low-temperature conditions

Pear fruit senescence under high-and low-temperature conditions has been reported to be mediated by microRNAs.Long non-coding RNAs(lncRNAs),which can function as competing endogenous RNAs that interact with microRNAs,may also be involved in temperature-affected fruit senescence.Based on the transcriptome and microRNA sequencings,in this study,3330 lncRNAs were isolated from Pyrus pyrifolia fruit.Of these lncRNAs,2060 and 537 were responsive to high-and low-temperature conditions,respectively.Of these differentially expressed lncRNAs,82 and 24 correlated to the mRNAs involved in fruit senescence under high-and low-temperature conditions,respectively.Moreover,three lncRNAs were predicted to be competing endogenous RNAs(ceRNAs)that interact with the microRNAs involved in fruit senescence,while one and two ceRNAs were involved in fruit senescence under high-and low-temperature conditions,respectively.A dual-luciferase assay showed that the interaction of an lncRNA with a microRNA disrupts the action of the microRNA on the expression of its target mRNA(s).Furthermore,four alternative splicing-derived lncRNAs interacted with miR172i homologies(Novel_88 and Novel_69)to relieve the repressed expression of their target and produce an miR172i precursor.Correlation analysis of microRNA expression suggested that Novel_69 is likely involved in the cleavage of the pre-miR172i hairpin to generate mature miR172i.Taken together,lncRNAs are involved in pear fruit senescence under high-or low-temperature conditions through ceRNAs and the production of microRNA.

Transcriptional profiles underlying the effects of salicylic acid on fruit ripening and senescence in pear(Pyrus pyrifolia Nakai)

Salicylic acid(SA) plays a pivotal role in delaying fruit ripening and senescence. However, little is known about its underlying mechanism of action. In this study, RNA sequencing was conducted to analyze and compare the transcriptome profiles of SA-treated and control pear fruits. We found a total of 159 and 419 genes differentially expressed between the SA-treated and control pear fruits after 12 and 24 h of treatment, respectively. Among these differentially expressed genes(DEGs), 125 genes were continuously differentially expressed at both treatment times, and they were identified as candidate genes that might be associated with SA-regulated fruit ripening and senescence. Bioinformatics analysis results showed that 125 DEGs were mainly associated with plant hormone biosynthesis and metabolism, cell wall metabolism and modification, antioxidant systems, and senescence-associated transcription factors. Additionally, the expression of several candidate DEGs in ripening and senescent pear fruits after SA treatments were further validated by quantitative real-time PCR(qRT-PCR). This study provides valuable information and enhances the understanding of the comprehensive mechanisms of SA-meditated pear fruit ripening and senescence.

Effects of high CO_2 treatment on green-ripening and peel senescence in banana and plantain fruits

Banana fruit（Musa,AAA group,cv.Brazil） peel fails to fully degreen but the pulp ripens normally at temperatures above24°C.This abnormal ripening,known as green-ripening,does not occur in plantains（Musa,ABB group,cv.Dajiao）.Based on the fact that un-completely yellowing was also observed for bananas in poorly ventilated atmospheres,in the present study,the effect of high CO2 with regular O2（21%） on banana ripening was investigated along with that on plantains at20℃.The results showed that high CO2 conferred different effects on the color changing of bananas and plantains.After6 d ripening in 20%CO2,plantains fully yellowed,while bananas retained high chlorophyll content and stayed green.In contrast to the differentiated color changing patterns,the patterns of the softening,starch degradation and soluble sugar accumulation in the pulp of 20%CO2 treated bananas and plantains displayed similarly as the patterns in the fruits ripening in regular air,indicating that the pulp ripening was not inhibited by 20%CO2,and the abnormal ripening of bananas in 20%CO2 can be considered as green ripening.Similar expression levels of chlorophyll degradation related genes,SGR,NYC and PaO,were detected in the peel of the control and treated fruits,indicating that the repressed degreening in 20%CO2treated bananas was not due to the down-regulation of the chlorophyll degradation related genes.Compared to the effect on plantains,20%CO2 treatment delayed the decline in the chlorophyll florescence（F√F_m values and in the mRNA levels of a gene coding small subunit of Rubisco（SSU）,and postponed the disruption of the ultrastructure of chloroplast in the peel tissue of bananas,indicating that the senescence of the green cells in the exocarp layer was delayed by 20%CO2,to more extent in bananas than in plantains.High CO2 reduced the ethylene production and the expression of the related biosynthesis gene,ACS,but elevated the respiration rates in both cultivars.The up-regulation of the expression of anaerobic respiration pathway genes,ADH and PDC,might be responsible for the subtle effect of high CO2 on the pulp ripening.Taken together,the atmosphere of high CO2 and regular O2,delayed the senescence of the green cells in the exocarp layer of the banana peel,but conferred no obvious inhibition on the pulp ripening,leading to a distinct green-ripening that was different from the phenomenon induced by high temperatures.