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Metabolomic and physiological analysis of bud differentiation in dense apple (Malus × domestica Borkh.) orchards following thinning and reshaping

ABSTRACT:

BACKGROUND: In China, a majority of apple orchards were initially planted under arboriculture. Noteworthy, as the trees age, there is an increase in the number of branches and the degree of orchard depression and deterioration of the canopy light conditions. This phenomenon results in in blockage of bud differentiation, causing yield decrease and overall deterioration in fruit quality. Intercropping is an effective strategy for addressing the issue of orchard depression. However, the impact of intercropping on bud differentiation remains unclear. This study thus analyzed the physiological and metabolic aspects of flower bud differentiation after thinning and reshaping an overcrowded orchard. METHODS: A series of physiological and metabolomic analyses were conducted on terminal flower bud samples collected during early (T1) and late (T2) bud differentiation following thinning and reshaping alongside their controls CK1 and CK2. RESULTS: The T1 group had significantly higher amounts of sucrose, glucose, fructose and sorbitol compared to its control. Metabolomic analyses yielded 845 metabolites. Among these metabolites, 72 were significantly different in CK1 vs T1. In contrast, 73 metabolites were different in CK2 vs T2, while 26 metabolites were common between the two comparison groups. CK1 vs T1 and CK2 vs T2 had 20 and 16 enhanced metabolic pathways, respectively. Bud differentiation was closely associated with the phenylpropane and flavonoid biosynthesis pathways. CONCLUSIONS: Thinning and reshaping of the apple orchard improved light penetration and increased the content of coumarins and eriodictyol, thereby promoting flower bud differentiation. This study provides new insights into how orchard management practices, such as thinning, pruning and shaping, promote flower bud development in apple trees.

INSTRUMENT(S): Liquid Chromatography MS - negative - reverse phase, Liquid Chromatography MS - positive - reverse phase

PROVIDER: MTBLS11951 | MetaboLights | 2025-01-20

REPOSITORIES: MetaboLights

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Project description:Experiments were carried out in 2008 on 8-year-old apple trees (cv. Golden Delicious/M9) trained with standard horticultural practices at the experimental farm of the IASMA (Trento, Italy). Populations of fruits with different abscission potentials (abscising fruitlets versus persisting fruitlets), were established as described by Dal Cin et al. (2005, 2007, 2009), Angeli et al. (2002), and other preliminary experiments (unpublished data). Briefly, the abscising population was made up of lateral fruitlets treated with benzylaminopurine (BA) at 200 ppm (commercial name Brancher-Dirado), when fruits had an average size of 13 mm (about 15 days after petal fall, DAPF). The population of central persisting fruitlets was generated by removing all the laterals from each cluster at petal fall, and leaving exclusively the central flower that had been hand-pollinated at full bloom with compatible pollen (cv. Stark Red). Samples of the two populations were collected at defined time points from groups of twenty homogeneous trees randomly distributed in the orchard in four blocks. Fruits were collected and categorised into three classes of size (class 1, smaller fruits; class 2, medium fruits; class 3, bigger fruits), two classes related to the position within the clusters (lateral versus central fruits), and two classes on the base of the treatment (BA-treated versus untreated fruits). Fruits of the intermediate size (class 2 fruits in Figure 1) were not considered for sampling and subsequent molecular analyses, and only fruits of the two more divergent size classes 1 and 3 were kept. This resulted in a combined categorisation of fruits into 4 classes: 1) untreated lateral fruitlets, 2) lateral fruitlets treated with BA, 3) untreated central fruitlets, and 4) central fruitlets treated with BA (Figure 1). Each class was further distinguished into the two size categories 1 (small fruits) and 3 (bigger fruits), for a total of eight experimental theses. Fruitlet shedding and ethylene evolution were monitored throughout the physiological drop from the beginning of the experiments up to 46 DAPF in all fruitlet classes, separately. Seed and cortex (including epidermis) samples were collected from all classes of fruitlets at 0 (T0), 1 (T1), 2 (T2), 4 (T3), 6 (T4), and 8 (T5) days after the BA treatment from control and treated trees, and according to their position within the clusters (central vs lateral) and size (small vs big), as described. The latter parameter was decided at each sampling date based upon the mean cross diameter of the whole population of lateral fruits, calculated over a sample of 100 fruits measured randomly on twenty trees. The small ones had a cross diameter below the mean - s.d. (standard deviation), whereas the big ones had a cross diameter above the mean + s.d. Lateral fruitlets were collected from intact clusters showing a clear hierarchy in terms of fruit size (i.e. with a clearly distinguishable central fruit, bigger than any lateral). Acronyms were ascribed to samples according to the following code: the first letter(s) describes fruitlet position within the cluster and the presence of BA treatment (L=untreated lateral, C=untreated central, LB=treated lateral, CB=treated central), then a digit to describe the size (1=small, 3=big), and finally a digit to described the time point (0=time of the treatment, 1=1 day after treatment, 2=2 days after treatment, 3=4 days after treatment, etc.). For example, LB32 is a lateral fruit, treated with BA, big sized, 2 days after treatment.<br>For microarray hybridizations, only samples of T0, T2, and T3 were used.

			Action	DRS
	neg_CK1.raw	Raw
	neg_CK2_1.raw	Raw
	neg_CK2_2.raw	Raw
	neg_CK2_3.raw	Raw
	neg_CK2_4.raw	Raw

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Metabolomic and physiological analysis of bud differentiation in dense apple (Malus × domestica Borkh.) orchards following thinning and reshaping

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