Metabolite annotations based on the integration of mass spectral information

A large number of metabolites are found in each plant, most of which have not yet been identified. Development of a methodology is required to deal systematically with unknown metabolites, and to elucidate their biological roles in an integrated 'omics' framework. Here we report the development of a 'metabolite annotation' procedure. The metabolite annotation is a process by which structures and functions are inferred for metabolites. Tomato ( Solanum lycopersicum cv. Micro-Tom) was used as a model for this study using LC-FTICR-MS. Collected mass spectral features, together with predicted molecular formulae and putative structures, were provided as metabolite annotations for 869 metabolites. Comparison with public databases suggests that 494 metabolites are novel. A grading system was introduced to describe the evidence supporting the annotations. Based on the comprehensive characterization of tomato fruit metabolites, we identified chemical building blocks that are frequently found in tomato fruit tissues, and predicted novel metabolic pathways for flavonoids and glycoalkaloids. These results demonstrate that metabolite annotation facilitates the systematic analysis of unknown metabolites and biological interpretation of their relationships, which provide a basis for integrating metabolite information into the system-level study of plant biology.     

Mutations in tomato 1-aminocyclopropane carboxylic acid synthase2 uncover its role in development beside fruit ripening

The role of ethylene in plant development is mostly inferred from its exogenous application. The usage of mutants affecting ethylene biosynthesis proffers a better alternative to decipher its role. In tomato (Solanum lycopersicum), 1-aminocyclopropane carboxylic acid synthase2 (ACS2) is a key enzyme regulating ripening-specific ethylene biosynthesis. We characterised two contrasting acs2 mutants; acs2-1 overproduces ethylene, has higher ACS activity, and has increased protein levels, while acs2-2 is an ethylene underproducer, displays lower ACS activity, and has lower protein levels than wild type. Consistent with high/low ethylene emission, the mutants show opposite phenotypes, physiological responses, and metabolomic profiles compared with the wild type. The acs2-1 mutant shows early seed germination, faster leaf senescence, and accelerated fruit ripening. Conversely, acs2-2 has delayed seed germination, slower leaf senescence, and prolonged fruit ripening. The phytohormone profiles of mutants were mostly opposite in the leaves and fruits. The faster/slower senescence of acs2-1/acs2-2 leaves correlated with the endogenous ethylene/zeatin ratio. The genetic analysis showed that the metabolite profiles of respective mutants co-segregated with the homozygous mutant progeny. Our results uncover that besides ripening, ACS2 participates in the vegetative and reproductive development of tomato. The distinct influence of ethylene on phytohormone profiles indicates the intertwining of ethylene action with other phytohormones in regulating plant development.     

Reproductive water supply is prioritized during drought in tomato

Reproductive success largely defines the fitness of plant species. Understanding how heat and drought affect plant reproduction is thus key to predicting future plant fitness under rising global temperatures. Recent work suggests reproductive tissues are highly vulnerable to water stress in perennial plants where reproductive sacrifice could preserve plant survival. However, most crop species are annuals where such a strategy would theoretically reduce fitness. We examined the reproductive strategy of tomato (Solanum lycopersicum var. Rheinlands Ruhm) to determine whether water supply to fruits is prioritized above vegetative tissues during drought. Using optical methods, we mapped xylem cavitation and tissue shrinkage in vegetative and reproductive organs during dehydration to determine the priority of water flow under acute water stress. Stems and peduncles of tomato showed significantly greater xylem cavitation resistance than leaves. This maintenance of intact water supply enabled tomato fruit to continue to expand during acute water stress, utilizing xylem water made available by tissue collapse and early cavitation of leaves. Here, tomato plants prioritize water supply to reproductive tissues, maintaining fruit development under drought conditions. These results emphasize the critical role of water transport in shaping life history and suggest a broad relevance of hydraulic prioritization in plant ecology.     

番茄花萼在蕾期至果实红熟期的发育形态学及其多样性研究

萼片是番茄花和果实的重要组成部分,影响着果实的商品性。本试验以40份不同类型番茄为材料,对萼片发育过程、形态描述指标及形态多样性进行研究。结果表明,番茄萼片属于宿萼,其形态随着花和果实的发育而变化,表现为闭合、微开、展开、收合、微开、变形到定形,定形萼片呈现包被、基平、上翘、直立和上卷5种状态;对定形萼片7个形态性状观测表明,变异系数由大到小依次为萼片卷曲度(84.43%)、萼片面积(45.54%)、上翘度(40.93%)、形状系数(36.05%)、萼片长(35.02%)、萼片厚(29.46%)和萼片宽(24.61%)。相关性分析表明,萼片长、萼片宽、萼片厚和萼片面积四者之间均极显著正相关;萼片形状系数(萼片长/萼片宽)与萼片长极显著正相关,而与萼片宽无显著相关关系;萼片卷曲度和上翘度之间极显著正相关,而与其他5个性状没有显著相关关系。主成分分析表明,由萼片长、萼片宽、萼片厚和萼片面积代表的大小性状(PC1),由形状系数代表的形状性状(PC2),以及由卷曲度和上翘度代表的形态性状(PC3)3个主成分对萼片形态变异的累计贡献率达87.50%;用形态性状指标对定形的5种类型萼片形态进行了描述。     

Effects of colonization of a bacterial endophyte,Azospirillum sp. B510, on disease resistance in tomato

A plant growth-promoting bacteria, Azospirillum sp. B510, isolated from rice, can enhance growth and yield and induce disease resistance against various types of diseases in rice. Because little is known about the interaction between other plant species and this strain, we have investigated the effect of its colonization on disease resistance in tomato plants. Treatment with this strain by soil-drenching method established endophytic colonization in root tissues in tomato plant. The endophytic colonization with this strain-induced disease resistance in tomato plant against bacterial leaf spot caused by Pseudomonas syringae pv. tomato and gray mold caused by Botrytis cinerea. In Azospirillum-treated plants, neither the accumulation of SA nor the expression of defense-related genes was observed. These indicate that endophytic colonization with Azospirillum sp. B510 is able to activate the innate immune system also in tomato, which does not seem to be systemic acquired resistance.     

Quantitative trait loci analysis of seed‐specialized metabolites reveals seed‐specific flavonols and differential regulation of glycoalkaloid content in tomato

Given the potential health benefits (and adverse effects), of polyphenolic and steroidal glycoalkaloids in the diet there is a growing interest in fully elucidating the genetic control of their levels in foodstuffs. Here we carried out profiling of the specialized metabolites in the seeds of the Solanum pennellii introgression lines identifying 338 putative metabolite quantitative trait loci (mQTL) for flavonoids, steroidal glycoalkaloids and further specialized metabolites. Two putative mQTL for flavonols and one for steroidal glycoalkaloids were cross‐validated by evaluation of the metabolite content of recombinants harboring smaller introgression in the corresponding QTL interval or by analysis of lines from an independently derived backcross inbred line population. The steroidal glycoalkaloid mQTL was localized to a chromosomal region spanning 14 genes, including a previously defined steroidal glycoalkaloid gene cluster. The flavonoid mQTL was further validated via the use of transient and stable overexpression of the Solyc12g098600 and Solyc12g096870 genes, which encode seed‐specific uridine 5′‐diphosphate‐glycosyltransferases. The results are discussed in the context of our understanding of the accumulation of polyphenols and steroidal glycoalkaloids, and how this knowledge may be incorporated into breeding strategies aimed at improving nutritional aspects of plants as well as in fortifying them against abiotic stress.     

Genetic dissection of the role of ethylene in regulating auxin-dependent lateral and adventitious root formation in tomato

In this study we investigated the role of ethylene in the formation of lateral and adventitious roots in tomato ( Solanum lycopersicum ) using mutants isolated for altered ethylene signaling and fruit ripening. Mutations that block ethylene responses and delay ripening – Nr ( Never ripe ), gr ( green ripe ), nor ( non ripening ), and rin ( ripening inhibitor ) – have enhanced lateral root formation. In contrast, the epi ( epinastic ) mutant, which has elevated ethylene and constitutive ethylene signaling in some tissues, or treatment with the ethylene precursor 1-aminocyclopropane carboxylic acid (ACC), reduces lateral root formation. Treatment with ACC inhibits the initiation and elongation of lateral roots, except in the Nr genotype. Root basipetal and acropetal indole-3-acetic acid (IAA) transport increase with ACC treatments or in the epi mutant, while in the Nr mutant there is less auxin transport than in the wild type and transport is insensitive to ACC. In contrast, the process of adventitious root formation shows the opposite response to ethylene, with ACC treatment and the epi mutation increasing adventitious root formation and the Nr mutation reducing the number of adventitious roots. In hypocotyls, ACC treatment negatively regulated IAA transport while the Nr mutant showed increased IAA transport in hypocotyls. Ethylene significantly reduces free IAA content in roots, but only subtly changes free IAA content in tomato hypocotyls. These results indicate a negative role for ethylene in lateral root formation and a positive role in adventitious root formation with modulation of auxin transport as a central point of ethylene–auxin crosstalk.     

Resistance to Candidatus Liberibacter solanacearum haplotype B in tomato landraces from Mexico

Candidatus Liberibacter solanacearum haplotype B (CLsoB) is an economically important pathogen of tomato (Solanum lycopersicum L.) crops in New Zealand and Central and North America. Currently, resistant cultivars of tomato are not available as a management tactic because breeding programmes lack sources of resistance. Therefore, the objective of this study was to identify sources of resistance in tomato to CLsoB. Forty-six landraces of tomato were collected from several states in Mexico and were inoculated with CLsoB using 20 infested adults of Bactericera cockerelli per plant. Two assays were done over two years under greenhouse conditions. In the first trial, landraces FC22 and FC44 showed a significantly higher proportion of resistant plants, less symptom severity and longer incubation time compared with the other forty-four landraces and two susceptible cultivars. In the second assay, resistance to CLsoB of the landraces FC22 and FC44 was confirmed because they had again significantly greater numbers of resistant plants, less symptom severity, relative lower CLsoB titers and longer incubation time relative to the other genotypes. All plants considered resistant from both assays had DNA of CLsoB. Results indicate that all resistant plants from these landraces are promising sources for the development of tomato cultivars with resistance to CLsoB.     

不同甘氨酸浓度对无菌水培番茄幼苗生长和氮代谢的影响

植物不但能吸收矿质氮(NH+4-N、NO-3-N),而且也能直接吸收有机态氮,如氨基酸、小分子蛋白质等.为探讨有机态氮浓度对番茄幼苗生长和氮代谢的影响,无菌水培条件下采用2个番茄品种(申粉918、沪樱932)设置4种不同浓度(0、1.5、3.0、6 0mmol·L-1)的甘氨酸态氮(Gly-N),研究了番茄幼苗干物质重、吸氮量、氮代谢相关产物和氮代谢关键酶活性.结果表明,无菌水培条件下,随营养液中Gly浓度的增加,番茄植株干物质重、总氮量、地上部和根系游离氨基酸、可溶性蛋白、地上部可溶性糖含量增加.与无氮对照相比,各处理均显著降低了番茄地上部淀粉含量(P<0.05),而Gly浓度对根系淀粉含量无显著影响.随营养液中Gly浓度的增加,番茄地上部和根系的硝酸还原酶(NR)、谷氨酸脱氢酶(NADH-GDH)、丙转氨酶(GPT)和谷草转氨酶(GOT)活性均提高.无氮对照的NR活性与1.5 mmol·L-1 Gly处理之间差异不显著,而与3.0 mmol·L-1和6.0 mmol·L-1 Gly两处理之间差异显著(P<0.05);1.5 mmol·L-1 Gly和3.0 mmol·L-1 Gly两个处理之间的地上部NADH-GDH、GPT和GOT活性差异不显著.Gly浓度与番茄植株干物质重、总氮量呈显著正相关(R2>0.905* *),这表明两个番茄品种均能直接吸收利用甘氨酸.沪樱932吸收Gly的能力显著大于申粉918(P<0.05).因此,Gly-N可以成为番茄生长的良好氮源,其生理效应受Gly浓度的影响;不同品种番茄对Gly的吸收利用能力不同.     

Effects of ethephon and norbornadiene on sterol and glycoalkaloid biosynthesis in potato tuber discs

The accumulation of glycoalkaloids that normally takes place in aerobically incubated potato ( Solanum tuberosum L.) tuber discs has been found to be inhibited by the ethylene-releasing substance ethephon. Using ethephon and the ethylene action inhibitor norborna-2,5-diene, the effect of ethylene on the synthesis of sterols and glycoalkaloids, which partly share their biosynthetic pathway, was investigated.
Control discs showed incorporation of (2-¹⁴C)mevalonic acid into free sterols, steryl esters, steryl glycosides and acylated steryl glycosides at 24 h, thereafter the radioactivity decreased in free sterols and steryl esters concomitant with the appearance of radioactivity in glycoalkaloids. Discs with ethephon additions contained more radioactivity in all sterol classes at all time-points, but no glycoalkaloids were formed.
The enzyme S-adenosyl- l -methionine:sterol C24 methyltransferase (SMT, EC 2. 1. 1. 41), located at one presumed branching point in the sterol and glycoalkaloid pathway, was characterized and found to exhibit similar characteristics as in other plants, but a lower specific activity. The activity of SMT increased in ageing tuber discs and this increase was further stimulated by ethephon, but inhibited by norborna-2,5-diene. The activity of the glycoalkaloid-specific enzyme UDP-glucose:solanidine glucosyltransferase (EC 2. 4. 1) also increased after slicing, but here ethephon additions counteracted the induction. The activity of the sterol-specific UDP-glucose:sterol glucosyltransferase (EC 2. 4. 1) was unaffected by either tuber slicing or ethephon additions.
The results indicate that ethylene stimulates sterol synthesis in wounded potato discs, and that the wound-induction of SMT is regulated by ethylene.     

A snapshot of the Chinese SOL Project

In 2003, the International Solanaceae Project (SOL) was initiated by an international consortium of ten countries including Korea, China, the United Kingdom, India, the Netherlands, France, Japan, Spain, Italy and the United States. The first major effort of the SOL aimed to produce a DNA sequence map for euchromatin regions of 12 chromosomes of tomato (Solanum lycopersicum) before 2010. Here we present an update on Chinese effort for sequencing the euchromatin region of chromosome 3.     

A multicomponent, elicitor-inducible cystatin complex in tomato, Solanum lycopersicum

We assessed the ability of the fungal elicitor arachidonic acid to induce cystatin genes in tomato (Solanum lycopersicum), using a cDNA expression library from arachidonate-treated leaves. The cDNAs of two novel cystatins were isolated, coding for an approx. 11-kDa protein, SlCYS10; and for a 23.6-kDa protein, SlCYS9, bearing an N-terminal signal peptide and a long, 11.5-kDa extension at the C terminus. Both genes were induced by arachidonate but not by methyl jasmonate, an inducer of the 88-kDa eight-unit cystatin, multicystatin, accumulated in the cytosol of leaf cells upon herbivory. A truncated form of SlCYS9, tSlCYS9, was produced by deletion of the C-terminal extension to assess the influence of this structural element on the cystatin moiety. As shown by kinetic and stability assays with recombinant variants expressed in Escherichia coli, deleting the extension influenced both the overall stability and inhibitory potency of SlCYS9 against cysteine proteases of herbivorous organisms. These findings provide evidence for a multicomponent elicitor-inducible cystatin complex in tomato, including at least 10 cystatin units produced via two metabolic routes.