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Mercy A. Olmstead, N. Suzanne Lang, Frank W. Ewers and Shirley A. Owens

Xylem vessel anatomy was examined in tissues surrounding the graft union of sweet cherry (Prunus avium L.) scion (stem) and nondwarfing, semi-dwarfing, or dwarfing rootstock (root) combinations, to characterize potential changes in anatomical features during the initial stages of graft union formation. Vessel element length, frequency, diameter, lumen area (LAV), and mean vessel hydraulic diameter (VDh) were examined in `Rainier' (P. avium) scion wood grafted onto nondwarfing `Colt' (P. pseudocerasus L. × P. avium) or `F 12/1' (P. avium) rootstock and semi-dwarfing `Gisela 6' [`Gi 6' (P. cerasus L. × P. canescens L.)], or dwarfing `Gisela 5' [`Gi 5' (P. cerasus × P. canescens)] rootstock systems in: heterograft combinations (commercial scion-rootstock combinations); homografts (scion and rootstock are the same genetic material); and reciprocal heterografts (rootstock tissue grafted onto scion tissue). Vessel element length was not affected by rootstock, but vessel frequency and lumen area in graft union tissues were smaller in `Rainier'/`Gi 5' (dwarfing combination) than in `Rainier'/`Colt' (nondwarfing combination). The heterograft combination of `Rainier'/`Gi 5' had smaller scion LAV, lower VDh and narrower vessels than homograft or reciprocal heterograft combinations. As callus differentiated into vascular elements, xylem rays in `Rainer'/`Gi 5' tended to develop at an acute angle to the longitudinal axis of the tree and there was an increase in nonfunctional phloem in `Rainer'/`Gi 5' compared to `Rainer'/`F 12/1'. Collectively, the data provides further evidence that a combination of smaller and fewer vessels in the scion and graft union, as well as irregular vessel orientations in the vascular tissue within dwarfing combinations could contribute to hydraulic resistance in the graft union resulting in reduced scion growth (dwarfing).

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Mercy A. Olmstead, N. Suzanne Lang, Gregory A. Lang, Frank W. Ewers and Shirley A. Owens

Dye transport through vascular pathways was examined in tissues surrounding the graft union of second-leaf, field-grown trees of `Lapins'/Gisela 5 (`Gi 5') (dwarfing) and `Lapins'/'Colt' (nondwarfing). Excavated, intact trees were allowed to take up xylemmobile dye via transpiration for 6 h before sectioning the tree into scion, graft union, and rootstock tissue. `Lapins'/'Gi 5' had a significantly larger stem cross-sectional area in the central graft union than did `Lapins'/'Colt'. Per unit cross section, dye transport of both `Lapins'/'Gi 5' and `Lapins'/'Colt' was significantly less in the graft union than in rootstock sections, with still less transported to scion tissues in `Lapins'/'Gi 5'. `Lapins'/'Gi 5' had a tendency to produce vascular elements oriented obliquely to the longitudinal axis of the tree. Dye was distributed more uniformly axially and radially across the graft union in `Lapins'/'Colt' than in `Lapins'/'Gi 5', with an apparent accumulation of dye in `Lapins'/'Gi 5' graft union. Xylem vessel diameters and vessel hydraulic diameters (VDh) were smaller overall in `Lapins'/'Gi 5' than in `Lapins'/'Colt'; however, graft unions in both had smaller VDh than did rootstock sections. These observations suggest reduced transport efficiency of xylem vessels in the graft union in `Lapins'/'Gi 5' may be due to smaller vessels, vascular abnormalities and/or increased amounts of callus and parenchyma tissue.

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Mercy A. Olmstead, N. Suzanne Lang*, Frank W. Ewers and Shirley A. Owens

Dwarfing rootstocks in sweet cherry (Prunus avium L.) have been planted worldwide. No single theory has emerged to answer why scion dwarfing occurs in fruit trees. This research examines the vascular pathway in a dwarfing cherry system to determine if physical limitations alter water transport as a possible dwarfing mechanism. Second-leaf `Lapins' trees grafted onto Gisela 5 (Gi5; dwarfing) and Colt (vigorous) rootstocks were field-grown in East Lansing, Mich. During maximum shoot elongation, trees were dug, placed into containers with safranin dye solution (0.1% w/v) for 6 hours and then removed for division (3-5 cm in length) based on location in scion, graft union, and rootstock tissue. Tissues were sectioned using a sliding microtome (120 μm) for examination with a laser confocal microscope (Zeiss LSM Pascal). Mean stem area and vessel diameter were measured; and mean hydraulic diameter was calculated for vessels in the area of dye translocation. Overall, Lapins/Gi5 stem area in the graft union was larger compared to Lapins/Colt; however dye translocation in Lapins/Gi5 was reduced compared to other tissues in the tree. Confocal microscopy indicated dye uptake through the grafted region was more uniformly distributed in Lapins/Colt than in Lapins/Gi5, with dye accumulation in areas of maximum translocation. Vessel diameter did not differ in these areas of translocation. However, in both combinations there was a reduction in mean hydraulic diameter of graft union sections, suggesting a reduction in vessel efficiency to translocate water in this region. Vascular system anomalies were more frequent in Lapins/Gi5, disrupting acropetal dye translocation. This suggests the greatest reduction in vascular transport is in Lapins/Gi5.

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Clifford W. Beninger, George L. Hosfield, Mark J. Bassett and Shirley Owens

Three common bean (Phaseolus vulgaris L.) seedcoat color (or glossiness) genotypes, differing from each other by a single substitution at a seedcoat locus, were analyzed for presence and concentration of three anthocyanins: delphinidin 3-O-glucoside, petunidin 3-O-glucoside, and malvidin 3-O-glucoside. The three anthocyanins were present in Florida common bean breeding line 5-593 (P C J G B V Asp), matte black (P C J G B V asp), and dark brown violet (P C J G b V Asp), but the amounts varied greatly depending on the genotype. Dark brown violet had 19% of the total anthocyanin content when compared to 5-593, whereas matte black had amounts intermediate between the two other genotypes. The B gene acts to regulate the production of precursors of anthocyanins in the seedcoat color pathway above the level of dihydrokaempferol formation, perhaps at the chalcone synthase or chalcone isomerase steps in the biosynthetic pathway. We hypothesize that B regulates simultaneously the flavonoid (color) and isoflavonoid (resistance) pathways. The I gene for resistance to bean common mosaic virus (BCMV) is known to be linked closely to B. It is therefore hypothesized that the I gene function may be to respond to BCMV infection by dramatically increasing (over a low constituitive level) production in the 5-dehydroxy isoflavonoid pathway, which leads to synthesis of the major phytoalexin, phaseollin, for resistance to BCMV. Alternatively, the B and I genes may be allelic. The Asp gene affects seedcoat glossiness by means of a structural change to the seedcoat. We demonstrate that Asp in the recessive condition (asp/asp) changes the size and shape of the palisade cells of the seedcoat epidermis, making them significantly smaller than either 5-593 or dark brown violet. Asp, therefore, limits the amounts of anthocyanins in the seedcoat by reducing the size of palisade cells.