Mechanisms Of SlMAPK11 Regulate Fruit Ripening Via Interacting With SlSnRK1 In Tomato

Tomato(Solanum lycopersicum)Fruit ripening is accompanied by changes in texture,flavor,color,nutrients,etc.,and finally reaches the edible state.Studying the fruit ripening mechanism is of great significance to improve fruit quality,advance / delayed marketing,and extend shelf life.In previous study,Dr.Song Jianwen,cloned the mitogen-activated protein kinase gene 11(SlMAPK11),a key gene that can regulate tomato seed dormancy,and found that it may also regulate tomato fruit ripening.In view of this,this experiment investigated the SlMAPK11 transgenic plants in the field many times,and found that SlMAPK11 can indeed affect tomato fruit ripening,and then verified whether its interaction gene SlSnRK1 also affects tomato fruit ripening.After determining the gene function,we further understand its mechanism of action through experiments such as yeast two-hybrid and Co-IP.It also implies that the dormancy of seeds may be regulated by a common gene during fruit development and seed formation.The main results of this study are as follows:1.SlMAPK11 delays tomato fruit ripening.CRISPR/Cas9 knockout of the SlMAPK11 gene results in fruit ripening earlier than the receptor background material,and overexpression of this gene causes fruit ripening later than the receptor background material.2.SlMAPK11 interaction gene SlSnRK1 can also delays tomato fruit ripening.CRISPR/Cas9 knockout of SlSnRK1 results in fruit ripening earlier than the receptor background material.3.SlMAPK11 and SlSnRK1 affect fruit ripening through ethylene signaling pathway.Compared to the receptor background material,knocking out SlMAPK11 and SlSnRK1 increased ethylene release,while overexpression of SlMAPK11 decreased ethylene release.4.Yeast two-hybrid and Co-IP experiments showed that SlSnRK1 interacted with RIN,an important transcription factor regulating tomato maturation,knocking out this gene increased RIN expression,while SlMAPK11 did not interact with RIN.The expression levels of SlMAPK11 and SlSnRK1 are not induced by exogenous ethylene(ethephon).5.CRISPR/Cas9 knockout of SlMAPK11 and SlSnRK1 reduced the sensitivity of green ripening fruits to exogenous ethylene(ethephon).6.The yeast two-hybrid experiment showed that SlMAPK11 interacted with ethylene signal transduction pathway genes ETR6 and EIN2.