HCI buffer (H) pH, 7.0, Continuous tried Tris Citrate-I (CTC-I) pH 8.0, Continuous Tris Citrate-II (CTC-II) pH 7.0, Tris-Borate-EDTA (TBE) pH 8.5 4 Enzymes tried Esterase (EST), Glucose-6-phosphate dehydrogenase (G6PDH), Malate dehydrogenase (MDH), Malic enzyme (ME), 6-phosphogluconate dehydrogenase (6PGDH), phosphoglucoisomerase (PGI), phosphoglucomutase (PGM), menadione reductase (MR), shikimic acid dehydrogenase (SKDH
Molecular markers: Allozymes.--The following 13 loci were interpreted: Acn, F16dp, G3pdh, Idh, Mdh-1, Mdh-2, Me, Pgi-2, Pgm, Skdh
, 6-pgd, Tpi-1 and Tpi-2.
Five isozyme systems were analysed: isocitrate dehydrogenase (IDH), phosphoglucoisomerase (PGI), shikimate dehydrogenase (SKDH), 6-phosphogluconate dehydrogenase (6-PGD) and phosphoglucomutase (PGM).
3) SKDH: 10 ml pH 8.0 Tris-HCl 1M, 50 mg shikimic acid, 15 mg MTT, 10 mg NADP, 5 mg PMS plus 90 ml distilled water.
The spike sample differed from the original population at ACP, GPI, and SKDH loci, and the bulk sample differed at AAT and SKDH (Table 1).
There were differences between the original population and the spike population at ACP, SKDH, and [TPI.sub.2], and between the original and bulk population at ACP, GPI, and [6PGD.sub.2].
globosa were used to verify putative isozyme loci for GOT, SkDH
, and LAP.
In addition, these bean lines (except PI 164778) carry the same fast [F.sub.1] allele for the SKDH locus.
Abbreviations: G X E, genotype x environment; SKDH, shikimate dehydrogenase.
Fe1 0.058 0.093(*) NS Fe2 -0.001 -0.001 NS Idh -0.006 0.010 NS Lap 0.186(***) 0.012 NS Me 0.165(***) 0.151(***) NS Pgi2 -0.022 -0.017 NS Pgm 0.040 -0.011 (*) Skdh
0.131(***) 0.044 (***) Tpi 0.059 0.028 NS
Results were obtained from 11 enzyme systems, including aldolase (ALD), fructose-bisphosphatase (FBP), glutamate oxaloacetate transaminase (GOT), hexokinase (HK), isocitrate dehydrogenase (IDH), leucine aminopeptidase (LAP), malate dehydrogenase (MDH), 6-phosphogluconate dehydrogenase (6PGDH), phosphoglucose isomerase (PGI), shikimate dehydrogenase (SkDH
), and triosephosphate isomerase (TPI).
These loci (and the number of alleles per locus) were: isocitrate dehydrogenase (Idh, 4), leucine amino peptidase (Lap, 3), phosphoglucomutase-1 and -2 (Pgm- 1, 2; Pgm-2, 3), shikimate dehydrogenase (Skdh
, 2), malate dehydrogenase-2 (Mdh-2, 2), 6-phosphogluconate dehydrogenase- 1 and -2 6-Pgd- 1, 2; 6-Pgd-2, 3), diaphorase-2 (Dia-2, 3), and triose phosphate isomerase-1 (Tpi-1, 2).