Dominance induction of fruitlet shedding in Malus × domestica (L. Borkh): molecular changes associated with polar auxin transport

BACKGROUND: Apple fruitlet abscission is induced by dominance, a process in which hormones such as auxin, cytokinins and strigolactone play a pivotal role. The response to these hormones is controlled by transcription regulators such as Aux/IAA and ARR, whereas auxin transport is controlled by influx and efflux carriers. RESULTS: Seven partial clones encoding auxin efflux carriers (MdPIN1_A, MdPIN1_B, MdPIN10_A, MdPIN10_B, MdPIN4, MdPIN7_A and MdPIN7_B), three encoding auxin influx carriers (MdLAX1, MdLAX2 and MdLAX3) and three encoding type A ARR cytokinin response regulators (MdARR3, MdARR4 and MdARR6) were isolated by the use of degenerate primers. The organization of the PIN multigene family in apple is closer to Medicago truncatula than to Arabidopsis thaliana. The genes are differentially expressed in diverse plant organs and at different developmental stages. MdPIN1 and MdPIN7 are largely more expressed than MdPIN10 and MdPIN4. During abscission, the transcription of these genes increased in the cortex whereas in the seed a sharp fall was observed. The expression of these genes was found to be at least partially controlled by ethylene and auxin. CONCLUSION: The ethylene burst preceding abscission of fruitlets may be responsible for the decrease in transcript level of MDPIN1, MDARR5 and MDIAA3 in seed. This situation modulates the status of the fruitlet and its fate by hampering the PAT from the seeds down through the abscission zone (AZ) and this brings about the shedding of the fruitlet

Flavour compounds in tomato fruits: identification of loci and potential pathways affecting volatile composition

The unique flavour of a tomato fruit is the sum of a complex interaction among sugars, acids, and a large set of volatile compounds. While it is generally acknowledged that the flavour of commercially produced tomatoes is inferior, the biochemical and genetic complexity of the trait has made breeding for improved flavour extremely difficult. The volatiles, in particular, present a major challenge for flavour improvement, being generated from a diverse set of lipid, amino acid, and carotenoid precursors. Very few genes controlling their biosynthesis have been identified. New quantitative trait loci (QTLs) that affect the volatile emissions of red-ripe fruits are described here. A population of introgression lines derived from a cross between the cultivated tomato Solanum lycopersicum and its wild relative, S. habrochaites, was characterized over multiple seasons and locations. A total of 30 QTLs affecting the emission of one or more volatiles were mapped. The data from this mapping project, combined with previously collected data on an IL population derived from a cross between S. lycopersicum and S. pennellii populations, were used to construct a correlational database. A metabolite tree derived from these data provides new insights into the pathways for the synthesis of several of these volatiles. One QTL is a novel locus affecting fruit carotenoid content on chromosome 2. Volatile emissions from this and other lines indicate that the linear and cyclic apocarotenoid volatiles are probably derived from separate carotenoid pools

MADS Box Transcript Amount is Affected by Ethylene during Abscission

Thinning of young fruit is an important agronomical practice to ensure the maximum economic production. This practice is based on the control of the natural self thinning process occurring during fruit development. At the early stages of fruit development (fruitlet), the vegetative part of the tree is competing with the reproductive part of the tree and within the fruit clusters the different fruitlets are competing with each other. As a result the least fit organ abscises, Ethylene and auxin play a central role in this event but the role of ethylene is not thoroughly understood because in other systems abscission occurs partly with ethylene independent processes. We have followed the early development of fruitlets and studied the transcription patterns of MADS-box and ethylene related transcripts. Furthermore, we verified that ethylene has an effect on the expression of some ethylene related and MADS BOX genes. We propose that the ethylene burst during abscission induction is similar to a stage 2 ethylene system and it is related to fruitlet growth by affecting transcript amount of MADS-BOXes which modulate seed development and cortex growth

Physiological, molecular and practical aspects of fruit abscission

Many fruit species bear an abundance of flowers which produce a surplus of fruit that the tree is unable to support. In anticipation of this the major fruit species developed an immature fruit physiological drop as a self regulatory mechanism. The physiological fruit drop leads to a reduction of the fruit number per tree, but this is still insufficient to guarantee, at harvest, fruits of good marketable size and maximum commercial yield. For this reason supplementary fruit thinning is performed to achieve the optimum fruit load. Taking into account the practical importance of the thinning operations, the elucidation of the molecular mechanisms underlying natural fruitlet abscission is important for improving fruit thinning techniques and setting up molecular strategies useful for screening new chemical thinners or for selecting self thinning cultivars. Fruitlet abscission is a highly co-ordinated event. It involves multiple changes in cell structure, metabolism and gene expression leading to cell separation occurring in an abscission zone (AZ). Anatomical, biochemical and molecular events associated with cell separation at fruit AZ level have been described. Ethylene and IAA are deeply involved in the regulation of abscission. The general interaction between the two hormones is manifested in their antagonistic relation when the IAA status controls sensitivity of the tissue to ethylene. On the other hand, ethylene is a potent inhibitor of IAA interfering with its polar transport. Recent advances on molecular aspects referring to ethylene biosynthesis and action, as well as IAA polar transport, in relation to apple and peach fruitlet abscission are discussed

Dominance induction of fruitlet shedding in <it>Malus × domestica </it>(L. Borkh): molecular changes associated with polar auxin transport

Abstract Background Apple fruitlet abscission is induced by dominance, a process in which hormones such as auxin, cytokinins and strigolactone play a pivotal role. The response to these hormones is controlled by transcription regulators such as Aux/IAA and ARR, whereas auxin transport is controlled by influx and efflux carriers. Results Seven partial clones encoding auxin efflux carriers (MdPIN1_A, MdPIN1_B, MdPIN10_A, MdPIN10_B, MdPIN4, MdPIN7_A and MdPIN7_B), three encoding auxin influx carriers (MdLAX1, MdLAX2 and MdLAX3) and three encoding type A ARR cytokinin response regulators (MdARR3, MdARR4 and MdARR6) were isolated by the use of degenerate primers. The organization of the PIN multigene family in apple is closer to Medicago truncatula than to Arabidopsis thaliana. The genes are differentially expressed in diverse plant organs and at different developmental stages. MdPIN1 and MdPIN7 are largely more expressed than MdPIN10 and MdPIN4. During abscission, the transcription of these genes increased in the cortex whereas in the seed a sharp fall was observed. The expression of these genes was found to be at least partially controlled by ethylene and auxin. Conclusion The ethylene burst preceding abscission of fruitlets may be responsible for the decrease in transcript level of MDPIN1, MDARR5 and MDIAA3 in seed. This situation modulates the status of the fruitlet and its fate by hampering the PAT from the seeds down through the abscission zone (AZ) and this brings about the shedding of the fruitlet.</p

Benzylaminopurine Application on Two Different Apple Cultivars (Malus domestica) Displays New and Unexpected Fruitlet Abscission Features

Background and Aims: It has been previously shown that abscission of apple fruitlets is preceded by an increase in ethylene evolution and in the amount of transcripts for 1-aminocyclopropane-1-carboxylate oxidase (ACO), an enzyme catalysing the final step in ethylene biosynthesis. These events are concomitant with shedding induction and chemical thinning. There are several thinners but their mode of action and efficacy is poorly understood. One of them is benzylaminopurine (BA), a cytokinin believed to act by enhancing vegetative activity and stressing the competition between shoots and fruitlets, thus leading to fruitlet shedding. Nevertheless, the specific mechanism of action of BA and the variable effect depending on apple cultivar (easy or difficult to thin) are poorly understood. Methods: Abscission, the amount of MdACO1 transcripts and other parameters were followed in immature apple fruits during the period of physiological drop. The cultivars studied were \u2018Golden Delicious\u2019 and the \u2018spur\u2019 type \u2018Red Delicious\u2019. BA was used as a thinning agent and was sprayed 14 d after petal fall (DAPF). Fruitlets were divided into central (C) and lateral (L) fruitlet populations. Key Results: Fruitlet size was significantly different between C and L fruitlets but it did not differ much between the populations within the same cultivar. C fruitlets were characterized by basal ethylene evolution while L fruitlets displayed an increase in hormone biosynthesis during abscission induction. Cluster composition evaluated by the L/C ratio differed in the two varieties, being almost unchanged throughout abscission induction in \u2018Golden Delicious\u2019 and progressively decreasing in \u2018Red Delicious\u2019. Shoot growth activity evaluated at the end of the season indicated a possible connection with both the ongoing abscission and BA application. MDACO1 transcripts were mainly detected in L fruitlets and the accumulation was related to total abscission in \u2018Golden Delicious\u2019, while in \u2018Red Delicious\u2019 expression was observed in both C and L fruitlets. Conclusions: BA probably exerts its thinning effect through vegetative growth. In the \u2018spur\u2019 type \u2018Red Delicious\u2019 the chemical is ineffective, probably due to a limited action on shoot growth due to genetic characteristics. The amount of MdACO1 transcripts in seeds is a good indicator of abscission

The ethylene biosynthetic and signal transduction pathways are differently affected by 1-MCP in apple and peach fruit

1-Methylcyclopropene (1-MCP) is an antagonist of ethylene for receptor binding sites and the effects of its application differ in relation to a number of factors including genotype and ripening physiology. Peach (Prunus persica L. Batsch cv. 'Summer Rich') and apple (Malus x domestica L. Borkh cv. 'Golden Delicious') fruits were incubated with 1-MCP (1 mu L l(-1)) for 24 h at 20 degrees C and respiration rate, ethylene production and fruit firmness, together with ACC synthase, ACC oxidase, ETR1, ERS1, and CTR1 gene expression patterns were assessed throughout the post-treatment phase. 1-MCP was confirmed to be effective in delaying ripening in apples while in peaches only a limited effect of the chemical was observed. A dramatic inhibition of ethylene biosynthesis and ACS gene expression was induced by 1-MCP in apples whereas no marked difference was observed in peaches between the two controls (in air and in sealed jars without 1-MCP) and the treated fruit. In apples, Md-ETR1 and Md-ERS1 gene expression was down-regulated by 1-MCP starting from the end of the treatment, while Md-CTR1 appeared negatively affected by the chemical at a later stage. Transcription of Pp-ETR1, Pp-ERS1 and Pp-CTR1 genes appeared unaffected in 1-MCP treated peaches. Differences in receptor transcript levels between control fruit maintained in air and those enclosed in sealed jars without 1-MCP may be due to an effect of CO2 that rapidly accumulates following incubation of ripening peaches. Results support the hypothesis that the different behaviour of peaches and apples in response to 1-MCP application might be related to differences in terms of ratio, expression patterns and/or turn-over of the ethylene receptors

RNA extraction from plant tissue - The use of calcium to precipitate contaminating pectic sugars

Several protocols and commercial kits are used for the extraction of nucleic acids from different plant tissues. Although there are several procedures available to remove sugars, which hinder the extraction of clean genomic DNA, there are few to assist with extraction of RNA. Those presently used include precipitations with ethylene glycol monobutyl ether or lithium chloride (LiCl), or centrifugation in cesium chloride (CsCl) gradients, but these generally either do not allow high recovery of RNA, are time consuming, rely on hazardous chemicals or need special equipment. Here we present the use of the simple cation, Ca2+, which has been tested and shown to be very efficient for the precipitation of high molecular weight pectic sugars during RNA extraction. Results are presented for different plant tissues, especially tissues of peach and apple fruits at varying ripening stages

Ethylene biosynthesis and perception in apple fruitlet abscission (Malus domestica L. Borck)

Abscission was studied in immature apple fruits (cv. Golden Delicious) during the physiological drop. Fruitlet populations, characterized by different abscission potential, were analysed. Non-abscising fruitlets (NAF) were obtained from central flowers borne in clusters where all the lateral flowers had been removed at bloom while abscising fruitlets (AF) were derived from lateral fruitlets of trees sprayed with benzylaminopurine (BAP) at 200 ppm, 17 d after petal fall (APF), when the fruit cross diameter was about 10-12 mm. Fruit shedding, monitored at the end of the June drop, was significantly different in the two populations, being less than 10%, and more than 90%, in NAF and AF, respectively. In AF, fruit drop peaked around 33 d after petal fall (APF) and was preceded by an increase in ethylene around 20 d APF. Transcript analysis was performed from 17-24 d APF, since preliminary experiments pointed out that major changes in expression of abscission related genes occurred within this period. Transcript accumulation of genes involved in ethylene biosynthesis (MdACS5B and MdACO) and action (MdERS1, MdETR1, and MdCTR1) was studied in the seed, cortex, peduncle, and abscission zone (AZ) of the two fruit populations. MdACS5B and MdACO transcripts accumulated along the experimental period in AF population, even though at a different magnitude, while ethylene evolution declined after peaking at day three. MdETR1, MdERS1, and MdCTR1 expression patterns depended on tissue and/or population. The ERS/ETR ratio was higher in AF than in NAF populations. Overall results pointed out that apple fruitlet drop is preceded by a stimulation of ethylene biosynthesis and a gain in sensitivity to the hormone

Fruit load and elevation affect ethylene biosynthesis andaction in apple fruit (Malus domestica L. Borkh) duringdevelopment, maturation and ripening

The influence of internal and external factors such as tree fruit load and elevation on ethylene biosynthesis and action was assessed during apple fruit development and ripening. Ethylene biosynthesis, as well as transcript accumulation of the hormone biosynthetic enzymes (MdACS1 and MdACO1), receptors (MdETR1 and MdERS1) and an element of the transduction pathway (MdCTR1), were evaluated in apples borne by trees with high (HL) and low (LL) fruit load. Orchards were located in two localities differing in elevation and season day degree sum. These parameters significantly affected the date of bloom and commercial harvest, and the length of the fruit developmental cycle. Trees from the low elevation (LE) bloomed and the fruit ripened earlier than those from the high elevation (HE), displaying also a shortened fruit developmental cycle. Dynamics of ethylene evolution was apparently not affected by elevation. The onset of ethylene evolution started 130 days after bloom (DAB) at both elevations. During early ripening, fruits from LL trees produced significantly more ethylene than those from HL trees. Expression analysis of MdACS1, MdACO1 and MdERS1 indicated that the transcript accumulation well correlated with ethylene evolution. MdCTR1 was expressed at constant level throughout fruit growth and development up to 130 DAB, thereafter, the transcript accumulation decreased up to commercial harvest, concurrently with the onset of ethylene evolution