The phospholipids and galactolipids of the pollen-coat and internal domains of two lines of Brassica napus, Wesroona and IXLIN, with different linoleic/linolenic acid ratios (18:2/18:3) have been characterized by normal phase silica high performance liquid chromatography and gas liquid chromatography. The polar lipids of the pollen-coat are similar to leaf lipids in the high proportion of galactolipids (almost 50%) and the fatty acids; 18:3, palmitic (16:0) and hexadecatrienoic (16:3). In contrast, the pollen internal domain, although rich in 18:3, 18:2 and 16:0, is composed primarily of phosphatidyl-choline, -ethanolamine, and -inositol whose 18:2/18:3 ratio is correlated with that of the seed generation. The difference between the two divergent 18:2/18:3 ratio lines is most evident in the internal domain phospholipids. The 18:2/18:3 ratio of the galactolipids of both pollen domains is not significantly effected by the line genotype. The results are interpreted in terms of the previously described 'prokaryotic' and 'eukaryotic' plant desaturation pathways (PG Roughan, CR Slack [1982] Annu Rev Plant Physiol 33: 97-132). We propose that the eukaryotic pathway is the major desaturation pathway providing polyunsaturated fatty acids to the haploid-specified internal domain in which the IXLIN genotype modifies the activity of the sn-2 linoleoyl phosphatidylcholine desaturase/s of the endoplasmic reticulum. In the diploid-specified pollen-coat, our evidence suggests that a combination of the prokaryotic and eukaryotic pathways contribute polyunsaturated fatty acids.
Macrocyclic nickel(II), copper(II) and cobalt(II) complexes (6), (7), (9), (11), (13), (17) and (19) have
been prepared from 2,2'-iminobisbenzaldehyde, the appropriate metal salt and the primary diamines
3,3'-iminobispropylamine (4), 2,2'iminobisethylamine (9, 2,2'-(iminodimethylene)bisaniline (8),
azobenzene-2,2'-diamine (10), 2,2'-iminobisaniline (12), 2-amino-N-(2'-aminophenyl)benzamide (16)
and 2-amino-N-(2'-aminobenzoyl)benzamide (18) respectively. The nickel(II) complex (15), zinc(11)
complex (21) and copper(II) complex (22) were also prepared by similar methods. 2,2'-(Benzene-
1,2-diyldiimino)bisbenzaldehyde (25) was prepared but did not undergo metal template reactions to
allow the formation of macrocyclic complexes.
Enzyme-linked immunosorbent assays (ELISA's) were developed for the determination of alkenyl glucosinolates (sinigrin, gluconapin) in crude extracts of seed meal and vegetative tissues of Indian mustard (Brassica juncea). The assays were constructed from two types of high-titer anti-sinigrin polyclonal antibodies derived from antisera raised against a conjugate of sinigrin hemisuccinate with bovine serum albumin. An ovalbumin-sinigrin hemisuccinate conjugate was used as the ELISA plate-coating antigen. In cross-reactivity studies, antisera showed high specificities toward the alkenyl side chain and the thiohydroximate moiety of the glucosinolate molecule and did not bind to desulfoglucosinolates. All antisera showed 100% cross-reactivity with gluconapin, and one type cross-reacted completely with progoitrin, the principal glucosinolate of rapeseed (Brassica napus). The most sensitive ELISA had a linear inhibition curve over 1 X mol of sinigrin, with 50% inhibition at 2.8 X lo-" mol per assay. Parallel analyses of mustard seed and cotyledon extracts using both an ELISA and a standard HPLC method gave values identical within the error limits of the two procedures. The ELISA is, however, more sensitive, less expensive, and much faster than any other glucosinolate assay of comparable specificity.to 1 X
Lipids were extracted from the diploid seed and haploid pollen of Brassica napus L. Two fractions of pollen lipids, namely the diploid-specified pollen-coat and the haploid-specified internal cytoplasmic lipids were obtained. Significant correlations exist between pollen and seed generations for linoleic (18∶2) and linolenic (18∶3) acids. In pollen internal storage lipids, the level of 18∶3 is positively correlated and the level of 18∶2 is negatively correlated with the level of 18∶3 in seed lipids. Evidence is presented that strongly supports the hypothesis that lipid biosynthesis occurs within the pollen and that synthesis is specified by haploid genes. These data support the concept of pollen selection, so that selecting among living pollen grains for superior individuals has potential as a new plant breeding tool for improving seed oil quality.
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