Abstract

Transgenic tomato (Solanum lycopersicum) plants in which either mitochondrial malate dehydrogenase or fumarase was antisense inhibited have previously been characterized to exhibit altered photosynthetic metabolism. Here, we demonstrate that these manipulations also resulted in differences in root growth, with both transgenics being characterized by a dramatic reduction of root dry matter deposition and respiratory activity but opposite changes with respect to root area. A range of physiological, molecular, and biochemical experiments were carried out in order to determine whether changes in root morphology were due to altered metabolism within the root itself, alterations in the nature of the transformants' root exudation, consequences of alteration in the efficiency of photoassimilate delivery to the root, or a combination of these factors. Grafting experiments in which the transformants were reciprocally grafted to wild-type controls suggested that root length and area were determined by the aerial part of the plant but that biomass was not. Despite the transgenic roots displaying alteration in the expression of phytohormone-associated genes, evaluation of the levels of the hormones themselves revealed that, with the exception of gibberellins, they were largely unaltered. When taken together, these combined experiments suggest that root biomass and growth are retarded by root-specific alterations in metabolism and gibberellin contents. These data are discussed in the context of current models of root growth and biomass partitioning.

Highlights

  • (Landschützetal, 1995;Carrareital, 2003;Yuietal, 2003;Nunes-Nesiet al., 2005,2007a;Lemaitret al., 2007;Studart-Guimarãeetsal.,2007);howevers,everal of thesestudieswererelativelycursoryD. espitethis fleaacstt,tthweyogsetnuedrieasllpycroorvriodbinorgcalteoeanreeavniodtehnecrew,fiotrhaant i(Lmapnodrstcahnürtotzleeeotftahl.,eT1C9A95c;yYculieientfallo.,w2e0rd0e3v) oerloipnmthene t coordinatioonfphotosynthetaincdrespiratormyetab

  • The carbohydratceontentofwild-typerootswas similar to thatpreviouslyreported(Carrariet al.,2005;Lou et al.,2007).ThesestudiesrevealedthattheFL lines werecharacterizebdydecreasedlevelsofGlc,Fru(significantsloy inlinesFLU and FL41),and Sue.Bycontrastt,hemMDH lineswereinvariantinall oftheseparameterws ithrespecttothewild type.,it is perhapsimportantto notethat neitherthetotalsolublecarbohydratceontennt orthe proteincontent(TableII) oftheFL lineswas significantlydifferenftromthoseofthewild type,withthe levelsofstarchbeingdecreasedin bothsetsoftransformant(ssignificantsloyinlinesFL41and all mMDH lines;TableII)

  • Rootmorphologyof6-week-oldgraftedscions.Graftintghe wild-typeleafstockonto transgenicrootstock(eithermMDH8 [dark graybars]or FL41 [lightgraybars])or the reciprocalgraftinlged to a restoratioinnbotbrootarea (A) and totalrootlength(B) tothoseofthe wild-typeself-grafte(Wd T:WT)control(dashed horizontaline).These parameterswere significantlayltered(reduced in mMDH8 and increasedin FL41)intheungrafte(dung)and self-grafte(sdelf)antisense lines(AL).,thegraftincgould notrestoretherootdrymass(C)

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Summary

Wild Type

The carbohydratceontentofwild-typerootswas similar to thatpreviouslyreported(Carrariet al.,2005;Lou et al.,2007).ThesestudiesrevealedthattheFL lines werecharacterizebdydecreasedlevelsofGlc (significantlyso inlinesFLU andFL63),Fru(significantsloy inlinesFLU and FL41),and Sue (significantolynlyin lineFL63).Bycontrastt,hemMDH lineswereinvariantinall oftheseparameterws ithrespecttothewild type.,it is perhapsimportantto notethat neitherthetotalsolublecarbohydratceontennt orthe proteincontent(TableII) oftheFL lineswas significantlydifferenftromthoseofthewild type,withthe levelsofstarchbeingdecreasedin bothsetsoftransformant(ssignificantsloyinlinesFL41and all mMDH lines;TableII). Redoxprofilingof thetransgenilcinesrevealeda significandtecreasesin reducedglutathionien theFL lines,whileoxidizedglutathionleevelswerereduced toapproximatel6y0%ofthatfoundinthewildtypein. Data arepresentedas means± seofsixindividualplantsperline.Values inboldfaceweredeterminedbythettestto be significantldyifferen(Pt< 0.05) fromthewild type.Values are presentedas μιτιοΙg"1 freshweight,exceptforprotein,which is μg g~1 freshweight

Glc Fru
Trp Tyramine Tyr Val
Despitetherelativepaucityofchangesin thelevels
AnalysisoftheInfluenceofModifiedShootMetabolism on RootFunctionin theTransgenics
DISCUSSION
Tyr Trehalose
MATERIALSAND METHODS
Root Respiration
StatisticaAl nalyses
Findings
Malate Metabolism and Root Development
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