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Raffinose and Stachyose Accumulation, Galactinol Synthase Expression, and Winter Injury of Contrasting Alfalfa Germplasms

^a Dep. of Agronomy, Purdue Univ., West Lafayette, IN 47907-1150 USA
^b Station de Recherches, Agriculture and Agri-Food Canada, 2560 Hochelaga Blvd., Sainte-Foy, QC, Canada G1V 2J3
^c Soils and Crops Research and Development Centre, Agriculture and Agri-Food Canada, 2560 Hochelaga Blvd., Sainte-Foy, QC, Canada G1V 2J3

View larger version (45K): [in a new window]   Fig. 1. Raffinose and stachyose accumulation in taproots of cold acclimating alfalfa cultivars differing in fall dormancy (FD) and winter hardiness. Norseman (FD = 1) is winter hardy, Lahontan (FD = 7) has intermediate winter hardiness, while CUF 101 (FD = 9) and Wadi Qurayat (FD = 11) are not winter hardy. Responses of the parent cultivar (O) and populations from this cultivar selected for greater (L) and less (H) fall dormancy are shown. Data were averaged over 2 yr. The least significant difference (LSD) at the 5% level of probability is provided in the top panel of each column. Experiment 3.

View larger version (33K): [in a new window]   Fig. 2. Cold acclimation-induced changes in sucrose concentration in taproots of alfalfa cultivars differing in fall dormancy (FD) and winter hardiness. Norseman (FD = 1) is winter hardy, Lahontan (FD = 7) has intermediate winter hardiness, while CUF 101 (FD = 9) and Wadi Qurayat (FD = 11) are not winter hardy. Responses of the parent cultivar (O) and populations from this cultivar selected for greater (L) and less (H) fall dormancy are shown. Data were averaged over two years. The least significant difference (LSD) at the 5% level of probability is provided. Experiment 3.

View larger version (79K): [in a new window]   Fig. 3. Northern blot analysis of galactinol synthase (msaCIF) transcript (1.4 kB) accumulation in plants of alfalfa. (A) Time course of galactinol synthase transcripts accumulation in crowns of the cold-tolerant cv. Apica during cold acclimation at 2°C and disappearance following transfer at 20°C. (B) Transcript level in leaves and crowns of the cold-sensitive CUF 101 (C) and the cold-tolerant cv. Apica (A) and Rambler (R) that were non-acclimated (N), cold-acclimated two weeks at 2°C (H2) or cold acclimated 2 wk at 2°C followed by 2 wk at -2°C (HF). Assessment of total RNA (10 µg) per lane was made by ethidium bromide staining. Experiment 1.

View larger version (25K): [in a new window]   Fig. 4. Relative level of expression of galactinol synthase gene (msaCIF) in crowns of the cold-tolerant cv. Apica and Rambler and the cold-sensitive cv. CUF 101. Plants were acclimated to natural temperature conditions in an unheated greenhouse as described in Castonguay and Nadeau (1998). Means ± SEM (n = 3). Experiment 2.

View larger version (26K): [in a new window]   Fig. 5. Relationship between rankings of total raffinose-family oligosaccharides (Total RFO, open symbols, broken line) and sucrose (closed symbols, solid line) in taproots in December and winter injury rankings of diverse alfalfa cultivars and germplasms. Data were averaged over two winters. Experiment 3.

View larger version (43K): [in a new window]   Fig. 6. Cold acclimation-induced changes in galactinol synthase (msaCIF) transcript (1.4 kB) accumulation in taproots of alfalfa cultivars differing in fall dormancy (FD) and winter hardiness. Norseman (N, FD = 1) is winter hardy, while CUF 101 (C, FD = 9) is not winter hardy. Transcript levels (1.4-kb band) for the parent cultivar (O) and populations from this cultivar selected for greater (L) and less (H) fall dormancy are shown. Roots were sampled at monthly intervals as plants cold acclimated. Equal loading of total RNA per lane (20 µg) was verified by ethidium bromide staining. Experiment 3.