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PubMed1998

Biokinetic studies in humans with stable isotopes as tracers. Part 1: A methodology for incorporation of trace metals into vegetables.

The metabolism and biokinetics of trace metals in humans can be successfully studied employing stable isotopes of the investigated elements as tracers. For the estimation of the bioavailability and the intestinal absorption from solid food, materials are required which have been intrinsically labelled with the chosen stable tracer, since the use of an extrinsic label may lead to erroneous results. Here a technique for producing intrinsically labelled vegetables is presented and optimized with regard to molybdenum, gadolinium and ruthenium, elements of interest in the field of radiation protection and/or nutrition. These feasibility studies were aimed to determine the most favourable conditions for the production of vegetables containing the selected tracers in amounts high enough to enable successful biokinetic studies in humans. In this optimization study the natural elements were used instead of the more expensive stable isotopes. Mo is readily absorbed both into cress (Lepidium sativum) and into french beans (Phaseolus vulg. var. nanus). Gd uptake into cress is moderate, while Ru may be easily and successfully incorporated only into sprouts of mung beans (Vigna radiata).

Biological Availability↗
PubMed1996

Cowpea flour: a potential ingredient in food products.

Cowpeas (Vigna unguiculata L. Walp) are an important grain legume in East and West African countries as well as in other developing countries. The U.S. is the only developed country producing large amounts of cowpeas. High protein (18 to 35%) and carbohydrate (50 to 65%) contents, together with an amino acid pattern complementary to that of cereal grains, make cowpeas potentially important to the human diet from a nutritional standpoint. Despite their potential as an inexpensive source of protein and energy, cowpeas are underutilized in the U.S. and other industrialized countries. A simple technology for preparing cowpea flour would stimulate increased consumption of this legume. This paper reviews the status of cowpea flour production technologies and uses of cowpea flour in various food products.

Africa, Eastern↗
PubMed2000

Mass transport of proform of a KDEL-tailed cysteine proteinase (SH-EP) to protein storage vacuoles by endoplasmic reticulum-derived vesicle is involved in protein mobilization in germinating seeds.

A vacuolar cysteine proteinase, designated SH-EP, is expressed in the cotyledon of germinated Vigna mungo seeds and is responsible for the degradation of storage proteins. SH-EP is a characteristic vacuolar proteinase possessing a COOH-terminal endoplasmic reticulum (ER) retention sequence, KDEL. In this work, immunocytochemical analysis of the cotyledon cells of germinated V. mungo seeds was performed using seven kinds of antibodies to identify the intracellular transport pathway of SH-EP from ER to protein storage vacuoles. A proform of SH-EP synthesized in ER accumulated at the edge or middle region of ER where the transport vesicle was formed. The vesicle containing a large amount of proSH-EP, termed KV, budded off from ER, bypassed the Golgi complex, and was sorted to protein storage vacuoles. This massive transport of SH-EP via KV was thought to mediate dynamic protein mobilization in the cotyledon cells of germinated seeds. We discuss the possibilities that the KDEL sequence of KDEL-tailed vacuolar cysteine proteinases function as an accumulation signal at ER, and that the mass transport of the proteinases by ER-derived KV-like vesicle is involved in the protein mobilization of plants.

Animals↗
PubMed1978

Localization of vicilin peptidohydrolase in the cotyledons of mung bean seedlings by immunofluorescence microscopy.

Vicilin peptidohydrolase, the protease that hydrolyzes the reserve proteins in the cotyledons of mung bean (Vigna radiata) seedlings, has been localized intracellularly by immunofluorescence microscopy using monospecific antibodies against the enzyme and rhodamine-coupled goat-anti-rabbit immunoglobulin G's. The enzyme can first be visualized after 3 days of seedling growth and is associated with small foci within the cytoplasm of the storage parenchyma cells farthest from the vascular bundles. On the 4th day of growth, the protease is also present in the numerous large protein bodies within these cells. Vicilin peptidohydrolase is known to be synthesized de novo starting on the 3rd day of growth. Our observations are therefore consistent with the interpretation that the enzyme is synthesized in the cytoplasm and subsequently transported to the protein bodies.

Cytoplasm↗
PubMed2002

Effects of elevated ultraviolet-B radiation on native and cultivated plants of southern Africa.

Seventeen herb, shrub and tree species of commercial and ecological importance in southern Africa were exposed at one location to ultraviolet-B (UV-B, 280-315 nm) radiation approx. 35 % above clear-sky background (control). The aims were to assess how UV-B affects canopy area, dry mass, and some biochemical and morphological properties of leaves, and to investigate whether differences between species are related to growth form of the plants. There was no pattern of response to UV-B related to growth form. Leaves of trees had altered chlorophyll a and b, carotenoid and flavonoid concentrations, but those of shrubs or herbs did not. Non-structural carbohydrates were unaffected. Smaller canopy areas and dry masses were observed under enhanced UV-B, but these were not statistically different among growth forms. There was a general insensitivity of species to elevated UV-B. Only five species had significantly altered leaf biochemical and morphological properties, canopy area and dry mass, the changes differing in magnitude. There was no consistent pattern of change in leaf thickness or biochemical composition with increased UV-B. Correlation analyses did not support the view that growth is less negatively affected in species with thick leaves or in those where leaf thickness increases, or in species with naturally high leaf flavonoid contents or that are able to synthesize additional flavonoids in response to UV-B enhancement. The analyses did not support the hypothesis that growth was inhibited by starch accumulation in leaves under elevated UV-B. However, changes in leaf shape did correlate with canopy area and dry mass, showing the importance of photomorphogenetic changes caused by UV-B which affect species' performance. We conclude that generalizations on plant sensitivity to UV-B based on growth form and functional type could be misleading, and that the great majority of economically important species of the region are likely to be insensitive to future UV-B increases. Notable exceptions include the Colophospermum mopane tree ecotypes chota and leslie and the arable annual Vigna unguiculata, both of which are traditional sources of livelihood to rural African populations and of importance to African industry and agriculture.

Plant Development↗
PubMed2005

Isolation and characterization of four ascorbate peroxidase cDNAs responsive to water deficit in cowpea leaves.

BACKGROUND AND AIMS: Abiotic stresses stimulate formation of active oxygen species in plant tissues. Among antioxidant mechanisms, H2O2 detoxication by ascorbate peroxidases (APX) plays an important role. Several APX isoforms exist in plant cells, and they have rarely been studied separately. The aim of this work was to study changes in cytosolic, peroxisomal, stromatic and thylakoid APX gene expression in response to progressive drought, rapid desiccation and application of exogenous abscisic acid in the leaves of cowpea (Vigna unguiculata) plants. METHODS: Two cowpea (V. unguiculata) cultivars, 'EPACE-1' which is drought-tolerant and '1183'which is drought-sensitive, were submitted to drought stress by withholding irrigation. Detached leaves were air-dried or treated with exogenous abscisic acid. APX cDNAs were isolated by PCR and cloned in plasmid vectors. Changes in gene expression were studied using reverse-transcription PCR. KEY RESULTS: Four new V. unguiculata cDNAs encoding putative cytosolic, peroxisomal and chloroplastic (stromatic and thylakoidal) APX were isolated and characterized. In response to the different treatments, higher increases in steady-state transcript levels of the cytoplasmic and peroxisomal APX genes were observed in '1183' compared with 'EPACE-1'. On the other hand, the expression of the chloroplastic APX genes was stimulated earlier in the tolerant cultivar when submitted to progressive drought. CONCLUSIONS: Water deficit induced differences in transcript accumulation of APX genes between the two cultivars that were related to their respective tolerance to drought. Chloroplastic APX genes responded early to progressive water deficit in the tolerant plant, suggesting a capacity to efficiently detoxify active oxygen species at their production site. The more sensitive '1183' was also able to respond to drought by activating its whole set of APX genes.

Abscisic Acid↗
PubMed2006

Water uptake by dry beans observed by micro-magnetic resonance imaging.

BACKGROUND AND AIMS: Water uptake by dry kidney beans (Phaseolus vulgaris 'Rajma') and adzuki beans (Vigna angularis) was traced using micro-magnetic resonance imaging in order to elucidate the channel of water entry, the manner of water delivery and the timing of swelling of the seeds. METHODS: Magnetic resonance images of beans absorbing water were continuously measured with the single-point imaging method for 16 h or 20 h at 15-min intervals. With this technique, it was possible to detect and visualize the location of water in the beans, at a low water content, in the initial stages of water entry. KEY RESULTS: Water was taken up through a specified tissue, the lens, near the hilum, and distributed primarily to the testa. When water reached the radicle, it began to be incorporated into cotyledons with considerable swelling of the seeds. Water uptake took place within a short time for kidney beans. The initial process of water entry was associated with mechanical vibration of the seed. Rapid hydration of the testa and the swelling of the cotyledons were then observed. Water was supplied to cotyledons through the adaxial epidermis. In contrast, it took a long time, approx. 7 h, to activate the water channel of the lens for adzuki beans which have a tightly fitting testa. Steeping of the testa was not uniform, which induced temporary slanting before enlargement of the seed. CONCLUSIONS: The activation of the lens as the sole water channel, the delivery of water to the radicle within the testa, the swelling of the cotyledons, and the further increment of water are physiologically different processes during imbibition, and were separated by locating water in various tissues and by analysing the time course of water uptake using magnetic resonance imaging with the single-point imaging method.

Cotyledon↗
PubMed2006

Glutathione reductase in leaves of cowpea: cloning of two cDNAs, expression and enzymatic activity under progressive drought stress, desiccation and abscisic acid treatment.

BACKGROUND AND AIMS: Reactive oxygen species are frequently produced when plants are exposed to abiotic stresses. Among the detoxication systems, two enzymes, ascorbate peroxidase and glutathione reductase (GR) play key roles. GR has also a central role in keeping the reduced glutathione pool during stress thus allowing the adjustments on the cellular redox reactions. The aim of this work was to study the variations in cytosolic and dual-targeted GR gene expression in the leaves of cowpea plants submitted to progressive drought, rapid desiccation and application of exogenous abscisic acid (ABA). METHODS: Two cowpea (Vigna unguiculata) cultivars, one drought-resistant ('EPACE-1'), the other drought-sensitive ('1183') were submitted to progressive drought stress by withholding irrigation. Cut-off leaves were air-dried or treated with exogenous ABA. Two GR cDNAs, one cytosolic, the other dual-targeted to chloroplasts and mitochondria were isolated by PCR and cloned in plasmid vectors. Reverse-transcription PCR was used to study the variations in GR gene expression. KEY RESULTS: Two new cDNAs encoding a putative dual-targeted and a cytosolic GR were cloned and sequenced from leaves of V. unguiculata. Drought stress induced an up-regulation of the expression of the cytosolic GR gene directly related to the intensity of the stress in both cultivars. The expression of dual-targeted GR was up-regulated by the drought treatment in the susceptible cultivar only. Under a fast desiccation, the '1183' cultivar responded later than the 'EPACE-1', although in 'EPACE-1' it was the cytosolic isoform which responded and in '1183' the dual-targeted one. Exogenous ABA enhanced significantly the activity and expression levels of GR in both cultivars after treatment for 24 h. CONCLUSIONS: These results demonstrate a noticeable activation in both cultivars of the antioxidant metabolism under a progressive water stress, which involves both GR genes in the case of the susceptible cultivar. Under a fast desiccation, the susceptible cultivar responded later than the resistant one, suggesting a weaker capacity of response versus the resistant one. Exogenous ABA probably acts on GR gene expression via a mediated signal transduction pathway.

Abscisic Acid↗
PubMed2005

In situ modification of herbivore-induced plant odors: a novel approach to study the attractiveness of volatile organic compounds to parasitic wasps.

Many parasitic wasps (parasitoids) exploit volatile organic compounds (VOCs) emitted by herbivore-infested plants in order to locate their hosts, but it remains largely unknown which specific compounds within the volatile blends elicit the attractiveness to parasitoids. One way of studying the importance of specific VOCs is to test the attractiveness of odor blends from which certain compounds have been emitted. We used this approach by testing the attraction of naive and experienced females of the two parasitoids Cotesia marginiventris and Microplitis rufiventris to partially altered volatile blends of maize seedlings (Zea mays var. Delprim) infested with Spodoptera littoralis larvae. Adsorbing filter tubes containing carbotrap-C or silica were installed in a four-arm olfactometer between the odor source vessels and the arms of the olfactometer. The blends breaking through were tested for chemical composition and attractiveness to the wasps. Carbotrap-C adsorbed most of the sesquiterpenes, but the breakthrough blend remained attractive to naive C. marginiventris females. Silica adsorbed only some of the more polar VOCs, but this essentially eliminated all attractiveness to naive C. marginiventris, implying that among the adsorbed compounds there are some that play key roles in the attraction. Unlike C. marginiventris, M. rufiventris was still attracted to the latter blend, showing that parasitoids with a comparable biology may employ different strategies in their use of plant-provided cues to locate hosts. Results from similar experiments with modified odor blends of caterpillar-infested cowpea (Vigna unguiculata) indicate that key VOCs in different plant species vary greatly in quality and/or quantity. Finally, experienced wasps were more strongly attracted to a specific blend after they perceived the blend while ovipositing in a host. Considering the high number of distinct adsorbing materials available today, this in situ modification of complex volatile blends provides a new and promising approach pinpointing on key attractants within these blends. Advantages and disadvantages compared to other approaches are discussed.

Adsorption↗
PubMed2002

Evolutionary re-organisation of a large operon in adzuki bean chloroplast DNA caused by inverted repeat movement.

We have sequenced two sections of chloroplast DNA from adzuki bean (Vigna angularis), containing the junctions between the inverted repeat (IR) and large single copy (LSC) regions of the genome. The gene order at both junctions is different from that described for other members of the legume family, such as Lotus japonicus and soybean. These differences have been attributed to an apparent 78-kb inversion that spans nearly the entire LSC region and which is present in adzuki and its close relative, the common bean. This 78-kb rearrangement broke the large S10 operon of ribosomal proteins into two smaller operons, one at each end of the LSC, without affecting the gene content of the genome. It disrupted the physical and transcriptional relationship between the six-gene rpl23-rpl14 cluster and the four-gene rps8-rpoA cluster that is conserved in most land plants. Analysis of the endpoints of the rearrangement indicates that it probably occurred by means of a two-step process of expansion and contraction of the IR and not by a 78-kb inversion.

Base Sequence↗
PubMed1997

Dispersal and parasitizing abilities of Eupelmus vuilleti (Hymenoptera: Eupelmidae) within a column of cowpea seeds.

Eupelmus vuilleti (Crawford) is an ectoparasitoid of the seed-eating beetle Bruchidius atrolineatus (Pic), which is an important pest of stored cowpea, Vigna unguiculata Walp, seeds in West Africa. Herein, we investigated the dispersal abilities of females within columns of seeds to assess the potential of E. vuilleti as a biological control agent of bruchids in cowpea granaries. The influence of host presence together with the 2 abiotic factors light and gravity on parasitoid movement and parasitization efficiency were analyzed. E. vuilleti females were able to travel through large seed masses and parasitize hosts located at the end of the seed column opposed to their introduction zone. Parasitoid movement was stimulated by light. E. vuilleti females exhibited a negative geotropism. Females introduced at the bottom of the seed column dispersed more and parasitized more hosts than females introduced at the top. Host presence had some influence on the dispersal of the parasitoids within the seed column at a host density of 10 infested seeds for 16,000-18,000 uninfested seeds. This depended on female introduction zone because gravity was the major factor influencing dispersal. The possible applications of these results for biological control of bruchids in cowpea granaries are discussed.

Animals↗
PubMed1999

Nymphal survival and movement of crawlers of Bemisia argentifolii (Homoptera: Aleyrodidae) on leaf surfaces of selected vegetables.

Bemisia argentifolii Bellows & Perring generally feeds and develops on the under leaf surface of most host plants, but in excess of 50% of the immature population may be found on the upper surface of some host species. This study determined the influence of leaf surface on survival of immature B. argentifolii and the likelihood of movement by the crawler (1st instar) from the upper (adaxial) leaf surface on selected vegetable hosts. Laboratory, tests were conducted on 5 vegetables: cantaloupe, Cucumis melo L.; collard, Brassica oleracea ssp. acephala de Condolle: cowpea, Vigna unguiculata (L.) Walpers ssp. unguiculata; pepper, Capsicum annuum L. ssp. annuum; and tomato, Lycopersicon esculentum Miller. Survival to the 1st instar was high (85-95%) on the upper leaf surface on all hosts. Survival to the adult stage was similar between whitefly nymphs on the upper and lower leaf surfaces within each host crop, except more survived on the lower surface of cowpea compared with the upper surface. Movement of the crawlers from the upper to the lower surface was high on pepper (approximately 80%), cantaloupe (approximately 55%), and cowpea (approximately 55%). Conversely, less movement to the lower surface was observed on collard (approximately 18%) and tomato (approximately 30%). The impetus to move from the upper surface appears primarily a response to feeding and tactile cues instead of responses to geotropic or phototropic stimuli.

Animals↗
PubMed2003

Biology of Cerotoma arcuatus (Coleoptera: Chrysomelidae) and field validation of a laboratory model for temperature requirements.

Cerotoma arcuatus Olivier is a polyphagous pest of legumes [soybean, Glycine max (L.); dry beans, Phaseolus vulgaris (L.); and cowpeas, Vigna unguiculata (L.)], all of which are considered important protein sources for humans and domestic animals. Studies on the biology and temperature requirements of C. arcuatus were made under laboratory and field (cage) conditions. In the laboratory, insects were reared on soybean plants in incubators held at 18, 20, 22, 25, 28, 30, or 32 degrees C, 70 +/- 10% RH, and a photoperiod of 14:10 (L:D) h. A degree-days (DD) model developed based on the incubator data were validated in the field based on air and soil temperatures. The duration of the egg, larva-to-adult, and egg-to-adult period was inversely correlated with the temperature within the range of 18-32 degrees C, with the highest viability found from 20 to 30 degrees C. The temperature threshold for development and the thermal constant for the egg phase and the larva-to-adult periods were 13.6 degrees C and 106.7 DD, 8.3 degrees C and 399.4 DD, and 10.7 degrees C and 489.0 DD, respectively. The DD model for the egg-to-adult period, calculated using constant temperatures in the laboratory, was found to be valid for populations of C. arcuatus in the field, based on the fluctuating air and soil temperatures, although air temperatures provided more precise predictions. These data provide support for the rational control of this pest through population predictions based on their temperature requirements.

Air↗
PubMed2000

Protein modification by Amadori and Maillard reactions during seed storage: roles of sugar hydrolysis and lipid peroxidation.

The non-enzymatic modifications of proteins through Amadori and Maillard reactions play an important role in the loss of seed viability during storage. In the present study, the contribution of sugar hydrolysis and lipid peroxidation to Amadori and Maillard reactions, and to seed deterioration was investigated in mung-bean (Vigna radiata Wilczek). The contents of glucose and lipid peroxidation products in seed axes increased significantly during storage. The accumulation of Amadori products in seed axes was correlated to the lipid peroxidation, whereas the accumulation of Maillard products was closely correlated to sugar hydrolysis. The rate of accumulation of Maillard products was not well correlated to the content of Amadori products in both seed axes and protein/glucose model system, reflecting the complex nature of Amadori and Maillard reactions. The content of Amadori products in seed axes increased during the early stages of seed ageing, whereas the content of Maillard products increased steadily during the entire period of storage. The accumulation of Maillard products in seed axes was associated with the decline of seed vigour. These data suggest that, during seed ageing, sugar hydrolysis and lipid peroxidation are coupled with non-enzymatic protein modification through Amadori and Maillard reactions.

Fabaceae↗
PubMed2002

Difference in delta(15)N signatures between nodulated roots and shoots of soybean is indicative of the contribution of symbiotic N(2) fixation to plant N.

Symbiotic N(2) fixation has a variable effect on the (15)N abundance of different parts of legumes. Increases in fixation result in (15)N enrichment of nodules, while decreases, in combination with an increased uptake of mineral N, result in (15)N depletion of the root system. The difference between soybean shoot and below-ground delta(15)N (Deltadelta(15)N=delta(15)N(shoot)-delta(15)N(belowground)) was assessed in hydroponic culture over a range of rates of supply of mineral N. The fractional contribution of N(2) fixation to N uptake (%Ndfa) was determined using the natural abundance (NA) technique with ryegrass as a reference plant. Deltadelta(15)N and %Ndfa were highly correlated, and the relationship was tested using the same soybean cultivar grown in pots in N-rich soil. Estimates of %Ndfa derived from the NA method and from the Deltadelta(15)N approach yielded near-identical values. A literature survey showed similar relationships between %Ndfa and Deltadelta(15)N with different growth stages of soybeans grown under glasshouse and field conditions, different cowpea (Vigna unguiculata) cultivars in the field, and tagasaste (Chamaecytisus proliferus) in hydroponic culture. Possible confounding and species-specific (either plant or Rhizobium spp.) influences are discussed. The difference in delta(15)N signatures between nodulated roots and shoots is confirmed as a robust means of quantifying %Ndfa: it is independent of reference plants and offers the possibility of estimating %Ndfa in soils where the isotope composition of mineral N closely matches that of atmospheric N(2).

Algorithms↗
PubMed2004

Molecular cloning and characterization of a cDNA encoding asparaginyl endopeptidase from sweet potato (Ipomoea batatas (L.) Lam) senescent leaves.

Asparaginyl endopeptidase is a cysteine endopeptidase that has strict substrate specificity toward the carboxyl side of asparagine residues, and is possibly involved in the post-translational processing of proproteins. In this report one full-length cDNA, SPAE, was isolated from senescent leaves of sweet potato (Ipomoea batatas (L.) Lam). SPAE contained 1479 nucleotides (492 amino acids) in the open reading frame, and exhibited high amino acid sequence homologies (c. 61-68%) with asparaginyl endopeptidases of Vicia sativa, Phaseolus vulgaris, Canavalia ensiformis, and Vigna mungo. SPAE probably encoded a putative precursor protein. Via cleavage of the N- and C-termini, it produced a mature protein containing 325 amino acids (from the 51st to the 375th amino acid residues), the conserved catalytic residues (the 173rd His and 215th Cys amino acid residues), and the putative N-glycosylation site (the 332nd Asn amino acid residue). Semi-quantitative RT-PCR and western blot hybridization showed that SPAE gene expression was enhanced significantly in natural senescent leaves and in dark- and ethephon-induced senescent leaves, but was much less in mature green leaves, stems, and roots. Phylogenic analysis showed that SPAE displayed close association with vacuolar processing enzymes (legumains/asparaginyl endopeptidases), which function via cleavage for proprotein maturation in the protein bodies during seed maturation and germination. In conclusion, sweet potato SPAE is probably a functional, senescence-associated gene and its mRNA and protein levels were significantly enhanced in natural and induced senescent leaves. The possible role and function of SPAE associated with bulk protein degradation and mobilization during leaf senescence were also discussed.

Amino Acid Sequence↗
PubMed1986

Structure of the trypsin-binding domain of Bowman-Birk type protease inhibitor and its interaction with trypsin.

The crystal structure of the complex formed by bovine trypsin and Bowman-Birk type protease inhibitor AB-I extracted from azuki beans (Vigna angularis) 'Takara' has been analyzed. The structure was solved by the application of the phase combination of single isomorphous phases and trypsin model phases, followed by phase improvement using the iterative Fourier technique. From the resulting electron density map, a three-dimensional atomic model of the trypsin binding domain of AB-I has been built. The peptide chain at the trypsin reactive site turns back sharply at Pro29 and forms a 9-residue ring (Cys24-Cys32). The 'front side' of this ring, consisting of the reactive site (Cys24-Met28), interacts with trypsin in a similar manner to other families of inhibitors and forms a stable complex, which seems to be maintained by the interactions with the 'back side' of this ring (Pro29-Cys34). The similar spatial arrangements of the 'back side' of this inhibitor and the 'secondary contact region' of the other inhibitors with respect to the reactive site suggest an important common role of these regions in exhibiting inhibitory activity.

Chemical Phenomena↗
PubMed1998

Cross-species amplification of soybean (Glycine max) simple sequence repeats (SSRs) within the genus and other legume genera: implications for the transferability of SSRs in plants.

We investigated the transferability of 31 soybean (Glycine max) simple sequence repeat (SSR) loci to wild congeners and to other legume genera. Up to 65% of the soybean primer pairs amplified SSRs within Glycine, but frequently, the SSRs were short and interrupted compared with those of soybeans. Nevertheless, 85% of the loci were polymorphic within G. clandestina. Cross-species amplification outside of the genus was much lower (3%-13%), with polymorphism restricted to one primer pair, AG81. AG81 amplified loci in Glycine, Kennedia, and Vigna (Phaseoleae), Vicia (Vicieae), Trifolium (Trifolieae), and Lupinus (Genisteae) within the Papilionoideae, and in Albizia within the Mimosoideae. The primer conservation at AG81 may be explained by its apparent proximity to the seryl-tRNA synthetase gene. Interspecific differences in allele size at AG81 loci reflected repeat length variation within the SSR region and indels in the flanking region. Alleles of identical size with different underlying sequences (size homoplasy) were observed. Our findings and the emerging patterns in other plant studies suggest that in contrast to animals, successful cross-species amplification of SSRs in plants is largely restricted to congeners or closely related genera. Because mutations in both the SSR region and the flanking region contribute to variation in allele size among species, knowledge of DNA sequence is essential before SSR loci can be meaningfully used to address applied and evolutionary questions.

Base Sequence↗