Impaired nodule function in Medicago polymorpha L. infected with alfalfa mosaic virus.

The effects of alfalfa mosaic virus (AMV) on growth, nodule formation and nodule function in the annual burr medic, Medicago polymorpha cv. Circle Valley, were investigated in glasshouse pot experiments. Systemically-infected plants from AMV-infected seed produced 21% less shoot dry weight and accumulated 24% less fixed nitrogen in shoots than healthy plants when harvested 53 d after germination. At day 75, infected plants showed similar shoot dry weight losses (22%), but the quantity of nitrogen fixed fell by only 15%. At day 53, soluble sugar, starch and bacteroid concentrations in nodules were unaffected by AMV infection, but nitrogenase specific activity was decreased by 25% and soluble amino acids by 20%. Although AMV infection resulted in no differences in the number of nodules formed in the first 11 d after germination or at any harvest thereafter, nodule mass was decreased by 23% for virus-infected plants at day 53. However this difference disappeared by day 75. Growth of AMV-infected plants was decreased probably because of impaired N2 fixation with nodule function affected rather than nodulation. Increased nodule mass relative to plant weight in virus-infected plants, seen at day 75, implied some degree of compensation for the limitation in N2 -fixing capacity. ELISAs for AMV antigen indicated that nodules were active sites of virus multiplication.

Tracing B-genome chromatin in Brassica napus x B. juncea interspecific progeny.

We used polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH) techniques to demonstrate the presence of Brassica B-genome chromosomes and putative B-genome introgressions in B. napus x B. juncea interspecific progeny. The B-genome--specific repeat sequence pBNBH35 was used to generate PCR products and FISH probes. The highest frequencies of viable progeny were obtained when B. napus was the maternal parent of the interspecific hybrid and the first backcross. B-genome--positive PCR assays were found in 34/51 fertile F2 progeny (67%), which was more than double the proportion found in fertile BC(1) progeny. Four B-genome--positive F(2)-derived families and 1 BC(1)-derived family were fixed or segregating for B. juncea morphology in the F(4) and BC(1)S(2), respectively, but in only 2 of these families did B. juncea-type plants exhibit B. juncea chromosome count (2n = 36) and typical B-genome FISH signals on 16 chromosomes. The remaining B. juncea-type plants had B. napus chromosome count (2n = 38) and no B-genome FISH signals, except for 1 exceptional F(4)-derived line that exhibited isolated and weak B-genome FISH signals on 11 chromosomes and typical A-genome FISH signals. B. juncea morphology was associated with B-genome--positive PCR signals but not necessarily with 16 intact B-genome chromosomes as detected by FISH. B-genome chromosomes tend to be eliminated during selfing or backcrossing after crossing B. juncea with B. napus, and selection of lines containing B-genome chromatin during early generations would be promoted by use of this B-genome repetitive marker.

A PCR based B-genome-specific marker in Brassica species.

Previous hybridisation studies showed that the repetitive DNA sequence pBNBH35 from Brassica nigra (genome BB, 2n=16) bound specifically to the B-genome and not to the A- or C-genomes of Brassica species. We amplified a sub-fragment of pBNBH35 from B. nigra by PCR, cloned and sequenced this sub-fragment, and confirmed that it was a 329-bp sub-fragment of pBNBH35. PCR and hybridisation techniques were used to confirm that the pBNBH35 sub-fragment was Brassica B-genome-specific. Fluorescence in situ hybridisation (FISH) in B. nigra, B. juncea (AABB, 2n=36) and B. napus (AACC, 2n=38) showed that the pBNBH35 sub-fragment was present on all eight Brassica B-genome chromosomes and absent from the A- and C-genome chromosomes. The pBNBH35 repeat was localised to the centromeric region of each B-genome chromosome. FISH clearly distinguished the B-genome chromosomes from the A-genome chromosomes in the amphidiploid species B. juncea. This is the first known report of a B-genome repetitive marker that is present on all B-genome chromosomes. It will be a useful tool for the detection of B chromosomes in interspecific hybrids and may prove useful for phylogenetic studies in Brassica species.