Leguminous plants establish endosymbiotic associations with both rhizobia (nitrogen fixation) and arbuscular mycorrhizal fungi (phosphate uptake). These associations involve controlled entry of the soil microsymbiont into the root and the coordinated differentiation of the respective partners to generate the appropriate exchange interfaces. As part of a study to evaluate analogies at the molecular level between these two plant-microbe interactions, we focused on genes from Medicago truncatula encoding putative cell wall repetitive proline-rich proteins (RPRPs) expressed during the early stages of root nodulation. Here we report that a novel RPRP-encoding gene, MtENOD11, is transcribed during preinfection and infection stages of nodulation in root and nodule tissues. By means of reverse transcription-polymerase chain reaction and a promoter-reporter gene strategy, we demonstrate that this gene is also expressed during root colonization by endomycorrhizal fungi in inner cortical cells containing recently formed arbuscules. In contrast, no activation of MtENOD11 is observed during root colonization by a nonsymbiotic, biotrophic Rhizoctonia fungal species. Analysis of transgenic Medicago spp. plants expressing pMtENOD11-gusA also revealed that this gene is transcribed in a variety of nonsymbiotic specialized cell types in the root, shoot, and developing seed, either sharing high secretion/metabolite exchange activity or subject to regulated modifications in cell shape. The potential role of early nodulins with atypical RPRP structures such as ENOD11 and ENOD12 in symbiotic and nonsymbiotic cellular contexts is discussed.

SummaryExtracellular lipo-oligosaccharides of Rhizobiurn, known as Nod factors, play a key role in the molocular signal exchange which leads to the specific nitrogen-fixing symbiotic association between the soil microbe and its host legume. The biological activity of Nod factors and their perception by the host plant during the earliest stages of the Rhizobiurrdlegume intel;sction have been studied using transgenic alfalfa carrying a fusion between the promoter of the early nodulin gene MtENOD12 and the ~-glucuronidase (GUS) reporter gene. Histochemical staining has shown that GUS accumulates specifically in the differentiating root epidermis, prior to and during root hair emergence, within 2-3 h following the addition of purified Rhizobium meliloti Nod factors. This precocious transcriptional activation of the MtENOD12 gene, reminiscent of that observed after inoculation with intact Rhizobium, implies that the Nod factor signal can be perceived at a developmental stage preceding root hair formation. GUS activity can be detected following treatment with a wide range of R. meliloti Nod factor concentrations down to 10 -13 M, and furthermore, this rapid response to the bacterial elicitor appears to be non-systemic. Significantly, MtENOD12-GUS expression is not observed after inoculation with a R. meliloti nodH mutant which synthesizes exclusively non-suIphated Nod factors. Indeed purified Nod factors which lack the sulphate substituent are approximately 1000-fold less active than their suIphated counterparts. Thus, the triggering of ENOD12 transcription in the alfalfa root epidermis is a rapid molecular response which is subject to the same host-specificity determinant (Nod factor suIphation) that governs the interaction between alfalfa and its bacterial symbiont.

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An extensive search for maize (Zea mays) genes involved in cell wall biosynthesis and assembly has been performed and 735 sequences have been centralized in a database, MAIZEWALL (http://www.polebio.scsv.ups-tlse.fr/MAIZEWALL). MAIZE-WALL contains a bioinformatic analysis for each entry and gene expression data that are accessible via a user-friendly interface. A maize cell wall macroarray composed of a gene-specific tag for each entry was also constructed to monitor global cell wall-related gene expression in different organs and during internode development. By using this macroarray, we identified sets of genes that exhibit organ and internode-stage preferential expression profiles. These data provide a comprehensive fingerprint of cell wall-related gene expression throughout the maize plant. Moreover, an in-depth examination of genes involved in lignin biosynthesis coupled to biochemical and cytological data from different organs and stages of internode development has also been undertaken. These results allow us to trace spatially and developmentally regulated, putative preferential routes of monolignol biosynthesis involving specific gene family members and suggest that, although all of the gene families of the currently accepted monolignol biosynthetic pathway are conserved in maize, there are subtle differences in family size and a high degree of complexity in spatial expression patterns. These differences are in keeping with the diversity of lignified cell types throughout the maize plant.

Transgenic maize (Zea mays) plants were generated with a construct harboring a maize caffeic acid O-methyltransferase (COMT) cDNA in the antisense (AS) orientation under the control of the maize Adh1 (alcohol dehydrogenase) promoter. Adh1-driven ␤-glucuronidase expression was localized in vascular tissues and lignifying sclerenchyma, indicating its suitability in transgenic experiments aimed at modifying lignin content and composition. One line of AS plants, COMT-AS, displayed a significant reduction in COMT activity (15%-30% residual activity) and barely detectable amounts of COMT protein as determined by western-blot analysis. In this line, transgenes were shown to be stably integrated in the genome and transmitted to the progeny. Biochemical analysis of COMT-AS showed: (a) a strong decrease in Klason lignin content at the flowering stage, (b) a decrease in syringyl units, (c) a lower p-coumaric acid content, and (d) the occurrence of unusual 5-OH guaiacyl units. These results are reminiscent of some characteristics already observed for the maize bm3 (brownmidrib3) mutant, as well as for COMT down-regulated dicots. However, as compared with bm3, COMT down-regulation in the COMT-AS line is less severe in that it is restricted to sclerenchyma cells. To our knowledge, this is the first time that an AS strategy has been applied to modify lignin biosynthesis in a grass species.Lignins are complex phenolic polymers present in all vascular plants. They provide rigidity to conducting xylem elements and fiber cells. Lignins are composed of C 6 C 3 units, principally p-hydroxyphenyl (H), guaiacyl (G), and syringyl (S) units, and are present in various proportions according to botanical, physiological, and cytological criteria (Lewis and Yamamoto, 1990). Throughout the plant kingdom, grass lignins appear to be particularly specialized because they contain not only H, G, and S units, but also additional p-hydroxycinnamic units such as p-coumaric and ferulic acids (Higuchi et al., 1967). Ferulic acid may be ester linked to wall polysaccharides and/or ether linked to G units, thereby forming bridges between lignins and polysaccharides (Jacquet et al., 1995), whereas p-coumaric acid is primarily ester linked to S lignin units in lignified walls (Ralph et al., 1994; Grabber et al., 1996).Lignification in dicotyledons has been extensively studied and most of the known lignin biosynthetic genes have been employed in genetic engineering experiments (for review, see Grima-Pettenati and Goffner, 1999). There are almost no molecular data on lignification in grasses (Collazo et al., 1992;McAlister et al., 1998;Pichon et al., 1998;Selman-Housein et al., 1999;Spangenberg et al., 2001). Although lignification in grass species is likely to share a high degree of similarity to other angiosperms, the aforementioned structural specificity of grass cell walls may also involve a certain degree of grass-specific regulatory mechanisms. An in-depth knowledge of lignification in Graminaeae is of utmost importance because lignins are one of the ...

-Forage plants are the basis of ruminant nutrition, and cell wall digestibility is the limiting factor of their feeding value. Cell wall digestibility is therefore "the" target for improving the feeding value of forage crops. Among annual forages, maize cropped for silage making is the most widely used, and much research in genetics, physiology and molecular biology of annual forages is devoted to maize. Sorghum, immature small grain cereals and straws of small grain cereals are also given to cattle. Some dicotyledons are or were also used, such as forage beets, kales, canola in temperate areas and amaranths in tropical and subtropical areas. Large genetic variation for cell wall digestibility was proved from both in vivo and in vitro experiments in numerous species. Among the regular maize hybrids (excluding brown-midrib ones), the NDF in vivo digestibility nearly doubled from 32.9 to 60.1%. Correlations between in vivo and in vitro estimates of cell wall digestibility were often close to 0.75, but in vitro estimates of cell wall digestibility significantly reduced the range of variation between genotypes. Despite lignin content is well known as the major factor making cell wall undigestible, breeding for a higher digestibility of plant only from a lignin content trait appeared impossible. Correlations between lignin content and cell wall digestibility were indeed greatly variable according to the genetic background. Moreover, enzymatic solubilities were excessively dependent on lignin, and correlation between in vivo estimates of cell wall digestibility and lignin content were always lower than correlation between in vitro estimates of cell wall digestibility and lignin content. Among brown-midrib genes, the bm3 mutant in maize, and the bmr12 (and possibly bmr18) mutant in sorghum, which are both altered in the COMT activity, appeared as the most efficient in cell wall digestibility improvement. Moreover, a great genetic variation in the efficiency of the maize bm3 gene for cell wall digestibility improvement was observed according to the genetic background, with a lower efficiency when the normal germplasm was of better cell wall digestibility. Efficient breeding maize and others annual forage plants demands a renewing of genetic resources. Because resources of interest in cell wall digestibility improvement could be of poor agronomic value, the best is likely to cell wall digestibility / ingestibility / annual forage / maize / genetic variation / breeding / lignin Résumé -Amélioration de la digestibilité des parois des fourrages annuels. Les plantes fourragères sont à la base de l'alimentation des ruminants, mais leur valeur nutritive est limitée par la digestibilité variable des parois végétales, dont l'amélioration est alors la cible essentielle des programmes de sé-lection. Parmi les fourrages annuels, le maïs est la plante la plus cultivée, et c'est sur cette espèce que porte l'essentiel de la recherche en génétique, physiologie et biologie moléculaire. Toutefois, le sorgho, les céréales à paill...

http://www.heliagene.org/HP, http://lipm-helianthus.toulouse.inra.fr/dokuwiki/doku.php?id=start, https://www.heliagene.org/PlasmoparaSpecies (soon available).

The expression of phenylpropanoid and related genes was investigated in bm1, bm2, bm3, and bm4 near-isogenic maize plants at the 4-5 leaf stage using a gene-specific cell wall macro-array. The bm3 mutant, which is mutated in the caffeic acid O-methyltransferase (COMT) gene, exhibited the lowest number of differentially expressed genes. Although no other phenylpropanoid gene had an altered expression, two distinct OMT and two cytochrome P450 genes were overexpressed suggesting the activation of alternative hydroxylation/methylation pathways. The bm1 mutant had the highest number of differentially expressed genes, all of which were under-expressed. Bm1 mutant plants were affected not only in cinnamyl alcohol dehydrogenase (bm1 related CAD) gene expression as expected, but also in the expression of other CAD/SAD gene family members and several regulatory genes including MYB, ARGONAUTE and HDZip. As originally believed, the bm1 mutation could be localized at the CAD locus, but more probably in a gene that regulates the expression of the CAD gene family. The profile of under-expressed genes in the bm2 mutant is nearly similar to that of bm1. These genes fell under several functional categories including phenylpropanoid metabolism, transport and trafficking, transcription factors and regulatory genes. As the bm2 mutant exhibited a lower guaiacyl (G) unit lignin content, the bm2 mutation could affect a regulatory gene involved, perhaps indirectly, in the regulation, conjugation or transport of coniferaldehyde, or the establishment of G-rich maize tissues. The pattern of gene expression in bm4 plants, characterized by the over-expression of phenylpropanoid and methylation genes, suggests that the bm4 mutation likely also affects a gene involved in the regulation of lignification.