Dry bean breeding programs have made significant advances in combating both abiotic and biotic stresses as well as improving plant architectural traits via selective breeding. Flooding can cause complete crop loss in dry bean. On the other hand, breeding for an upright architecture in dry bean has been a breeding target in several programs. However, the stem cell wall components underlying this change have yet to be studied. This research focused on analyzing the cell wall components that might be involved in dry bean architecture as well as pre-germination flooding tolerance in dry bean. For the plant architecture study, two significant genomic regions were identified on Pv07 and Pv08 associated with lignin accumulation in dry bean. For the pre-germination flooding study, one unpigmented seed coat genotype (Verano) and three pigmented seed coat genotypes (Indeterminate Jamaica Red, Durango, and Midnight) had germination rates similar to that of the tolerant check.

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<i>Pinus taeda</i> L. suspension culture cells were used to develop a model system to study the process of lignification occurring during the early stages of cell wall formation and maturation. Chemical, biochemical and histochemical analyses of the <i>P. taeda</i> suspension cultures grown with 2,4-dichlorophenoxyacetic acid (2,4-D) as the growth regulator did not provide conclusive evidence for lignin deposition. On the other hand, cultures in which 2,4-D was substituted with α-naphthaleneacetic acid (NAA) were shown to lignify. During this induction of lignification, limited cell wall thickening occurred since transmission electron microscopy of the 2,4-D grown cells showed only primary walls while the average cell wall thickness of the NAA-grown cells was consistent with secondary (S₁) layer formation. Despite the possibility of only limited lignin deposition in the 2,4-0 grown cells, secondary metabolism had occurred as evidenced by reversed-phase and chiral chromatographic separations which revealed the ability of these cells to produce enantiomerically pure (-)-matairesinol. Administrations of [1-¹³C], [2-¹³C ] and [3-¹³C ] specifically labeled phenylalanines to the <i>P. taeda</i> suspension cultures in medium containing NAA allowed the determination of lignin bonding patterns <i>in situ</i> by solid-state ¹³C NMR spectroscopy of the resulting ¹³C enriched cells. Aqueous and organic solvent extractions and protease treatment yielded ¹³C enriched cell walls for solid-state ¹³C NMR spectroscopic analyses of the cell wall bound lignin component. Subsequently, an isolated lignin derivative from these cell walls was analyzed by solution-state ¹³C NMR spectroscopy and verified the assignments made in the solid-state. Accordingly, the above experiments represent the first demonstration of lignin bonding patterns <i>in situ</i> in a <i>Pinus</i> species as well as a suspension culture. This culture system possesses great potential as a model to thoroughly study the early stages of lignification. / Ph. D.

京都大学 / 新制・課程博士 / 博士(農学) / 甲第24670号 / 農博第2553号 / 新制||農||1099(附属図書館) / 学位論文||R5||N5451(農学部図書室) / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 梅澤 俊明, 教授 矢﨑 一史, 教授 森 直樹 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

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Changes in cell wall composition and in vitro degradation of corn stover fractions (leaf, upper stem and lower stem) with advancing maturity and in response to NH; treatment were determined, and possible inhibitory mechanisms of lignin were evaluated. With advancing maturity, IVDMD decreased (P<.001), associated with decreases (P<.001) in CP and water soluble carbohydrates (WSC), and increases (P<.001) in NDF and ADF. The IVDMD of leaf was higher (P<.001) than of stems, associated with higher CP, hemicellulose:cellulose, and arabinan:xylan, and lower lignin methoxyl content. A hypothesis of formation of reactive quinone methide intermediates from lignin during rumen fermentation was tested in vitro by incubating corn stover fractions with S-containing reducing agents. Sulphur incorporation into residual fiber occurred (P<.05), indicative of nucleophilic addition to quinone methide intermediates. Degradation of NDF was highly correlated with lignin methoxyl content. The impact of lignin on cellulose degradation was studied using lignocellulosic hydrogels, in which hydroxypropylated or unmodified hardwood lignin was blended with cellulose. In vitro cellulose degradation of lignocellulose blends was higher (P<.01) than of control. Addition of lignin at incubation depressed (P<.01) cellulose degradation. Hydroxypropylation enhanced (P<.001) the increase in cellulose degradation with lignin blending, and reduced (P<.001) the inhibitory effect of lignin addition at incubation. Treatment of drought-stressed corn stover with 3% aqueous NH₃ decreased (P<.05) NDF, compared to isonitrogenous NH₃ addition and control, associated with solubilization of hemicellulose. Esterified phenolic acids were released (P<.05) by NH₃ treatment in upper stem. The IVDMD and NDF degradation increased (P <.001) after ammoniation, with higher (P<.05) values for NH₃ treatment than NH₃ added in leaf. The in vitro response to ammoniation of fractions of drought-stressed and non-drought stressed corn stover harvested in subsequent years was compared, using N-sufficient and N-limiting buffers. Response was highest (P<.001) for non-drought stressed stover fractions, and in N-limiting medium. Response appeared to be affected by high concentration of WSC in lower stalks of drought-stressed stover. / Ph. D.