Heterotrimeric G proteins have been previously linked to plant defense; however a role for the Gbetagamma dimer in defense signaling has not been described to date. Using available Arabidopsis (Arabidopsis thaliana) mutants lacking functional Galpha or Gbeta subunits, we show that defense against the necrotrophic pathogens Alternaria brassicicola and Fusarium oxysporum is impaired in Gbeta-deficient mutants while Galpha-deficient mutants show slightly increased resistance compared to wild-type Columbia ecotype plants. In contrast, responses to virulent (DC3000) and avirulent (JL1065) strains of Pseudomonas syringae appear to be independent of heterotrimeric G proteins. The induction of a number of defense-related genes in Gbeta-deficient mutants were severely reduced in response to A. brassicicola infection. In addition, Gbeta-deficient mutants exhibit decreased sensitivity to a number of methyl jasmonate-induced responses such as induction of the plant defensin gene PDF1.2, inhibition of root elongation, seed germination, and growth of plants in sublethal concentrations of methyl jasmonate. In all cases, the behavior of the Galpha-deficient mutants is coherent with the classic heterotrimeric mechanism of action, indicating that jasmonic acid signaling is influenced by the Gbetagamma functional subunit but not by Galpha. We hypothesize that Gbetagamma acts as a direct or indirect enhancer of the jasmonate signaling pathway in plants.

Medicago sativa, alfalfa or lucerne, and M. arborea were considered reproductively isolated until recently. Then, in 2003, an alfalfa genotype was identified that produced a few seeds and progeny with hybrid traits after a large number of pollinations by M. arborea. A derivative of this alfalfa genotype also produced a low frequency of progeny with hybrid traits. Thus, the hybridization barrier was weakened by selection of seed parents. Hybrids from both events expressed traits from M. arborea and M. arborea-specific DNA bands, although more of the M. sativa genome was retained, based on the DNA results. Thus, there was chromatin elimination during embryogenesis, resulting in partial hybrids (hereafter hybrids). However, more than 30 hybrids with an array of M. arborea traits have been obtained thus far, and research continues on the nature of the hybrids. Traits have been genetically transmitted in crosses, and selected traits are in use for alfalfa breeding. This paper reviews the first hybrids and then focuses on further weakening of the hybridization barrier with the discovery of a more efficient hybridizer derived from crossing Medicago sativa subspecies, sativa, coerulea and falcata. This genotype was found to have reproductive abnormalities associated with its complex subspecies origin that are best described as hybrid breakdown. In effect, this subspecies derivative is a bridge-cross parent that consistently produces hybrids. Reproductive abnormalities in the bridge-cross parent are reported and discussed.

Five asymmetric hybrid plants were obtained between Medicago sativa (2n = 4x = 32) and Medicago arborea (2n = 4x = 32) through sexual reproduction and the use of a cytoplasmically male sterile M. sativa genotype. Over 2,000 pollinations were made to obtain these hybrids. Amplified fragment length polymorphism (AFLP) analysis showed that in the most studied hybrid (WA2273), 4% of the bands unique to the M. arborea parent were present, versus 72% for the unique M. sativa bands. This suggests that only a single M. arborea chromosome or chromosome parts has been transferred. WA2273 had 7% of AFLP bands which were not present in either parent, which is suggestive of chromosome rearrangements as would be expected if only chromosome parts or a single part had been transferred from M. arborea. Phenotypic evidence for hybridity was obtained for pod coiling (1.4 coils in WA2273 versus three coils in the M. sativa parent and its self and testcross populations, and one coil in M. arborea), and Colletotrichum trifolii race 2 resistance (transferred from the resistant M. arborea parent, as the M. sativa parent and the self populations were highly susceptible). The hybrids were self sterile, but were female fertile to a high level when crossed with 4x, but not 2x, M. sativa, indicating they were at or near 4x. Both the pod coiling trait and anthracnose resistance segregated in the progeny of testcrosses between WA2273 and M. sativa. The work demonstrates that agronomically useful traits can be introgressed into M. sativa from M. arborea by use of male sterile M. sativa and sexual reproduction.

Testcrosses were made with novel sources of lucerne germplasm. These were evaluated in the field in a subtropical environment to identify the lines which produced the highest yielding hybrids as a guide to future breeding efforts. The novel sources were derivatives of Medicago sativa × M. arborea partial (asymmetric) hybrids (termed sac) and very highly winter-active Omani landraces of M. sativa. As testers, 2 lines were used; a Colletotrichum trifolii race 2 resistant selection from the group 9 Australian-bred and adapted cultivar PacL 901 (selection hereafter termed 901) and the Omani landrace, Oman 2, collected at 17°N latitude, from Salalah, Oman. In the row experiment, substantial and significantly positive tester parent heterosis for overall yield (sum of 13 harvests) was observed in all of the sac × Oman 2 testcrosses, with the mean performance of the 11 testcrosses (1839 g/m row) significantly (P < 0.05) exceeding the mean performance of the sac × 901 testcrosses (1703 g/m row) evaluated. Where 901 was used as the tester, heterosis values relative to the tester for the same sac lines were negative for all testcrosses with 8 of the testcrosses being significantly negative. For the Omani landrace × 901 testcrosses, positive and negative heterosis values for total yield relative to the tester were observed, but none were significantly different from zero. The 901 tester yielded significantly (P < 0.05) more per se than the Oman 2 tester (1956 v. 1470 g/m row), although in an adjacent sward experiment Oman 2 yielded comparably to most of the standard commercial cultivars. The potential of the novel germplasm in the subtropics was verified in sward experiments with synthetics and/or strain crosses with yield increases of up to 42% over the benchmark synthetic Sequel. Further improvements can be expected following selection for disease and pest resistance within the lines and in the case of Oman 2 and sac, converging to maximise complementary gene action.