Growth, sucrose synthase, and invertase activities of developing Phaseolus vulgaris L. fruits

Activities of the sucrose-cleaving enzymes, acid and neutral invertase and sucrose synthase, were measured in pods and seeds of developing snap bean (Phaseolus vulgaris L.) fruits, and compared with ¹⁴C-import, elongation and dry weight accumulation. During the first 10 d post-anthesis, pods elongated rapidly with pod dry weight increase lagging behind by several days. The temporal patterns of acid invertase activity and import coincided closely during the first part of pod development, consonant with a central role for this enzyme in converting imported sucrose during pod elongation and early dry weight accumulation. Later, sucrose synthase became the predominant enzyme of dry weight accumulation and was possibly associated with the development of phloem in pod walls. Sucrose synthase activity in seeds showed two peaks, corresponding to two phases of rapid import and dry weight accumulation; hence, sucrose synthase was associated with seed sink growth. Acid invertase activities in seeds were low and did not show a noticeable relationship with import or growth. All neutral invertase activities, during pod and seed development, were too low for it to have a dominant role in sucrose cleavage. Changes in activities of certain sucrose-cleaving enzymes appear to be correlated with certain sink functions, including import, storage of reserves, and biosynthetic activities. The data supports the association of specific sucrose-cleaving enzymes with the specific processes that occur in the developing pods and seeds of snap bean fruits; for example, acid invertase with pod elongation and sucrose synthase with fruit dry matter accumulation.     

A model to predict seed number per pod from early pod growth rate in pea (Pisum sativum L.)

Seed number per pod in pea is variable. After fertilization,any number of ovules may abort in the pod. This paper describesan analysis of the relationship between pod growth and seedabortion rate. Pod dry weight was highly correlated to pod lengthbefore the final stage in seed abortion. By measuring pod lengthduring the period of seed formation and counting the seeds inthe same pods at the end of this period, we show that seed numberper pod was correlated with early pod elongation. From thesedata we propose and test a model for predicting seed numberper pod from early pod elongation rates. Key words: Pea, pod length, pod growth, seed number, modelling     

Application of a probabilistic model for analysing the abortion of seeds and pods in winter oilseed rape (Brassica napus)

In winter oilseed rape (WOSR), only a subset of ovules can develop into seeds in the majority of pods. Any difficulty during the process of seed production may result in seed or pod abortion. This study aimed to reproduce the process of seed development in WOSR based on a limited number of parameters. As a result of the complexity of the developmental patterns of WOSR, it is challenging to identify the roles of various factors that influence seed production using an experimental approach. Here, we present a stochastic probabilistic model of seed development. The generalised least squares method was implemented to estimate the model parameters using the experimental data. Experiments were done in Grignon (France) in 2008 and 2009. The variations in the parameters were analysed according to the following four factors: year, pod rank, inflorescence position and ramification‐clipping treatment. The year had no effect on the number of ovules per ovary (μ) and the probability of seed viability (p). The proportion of effective pollen grains (k) significantly decreased with pod rank at the end of the main stem. Inflorescence position influenced the number of ovules per ovary (μ: 30.8–33.8 from top to bottom) and the parameter k. The mean number of seeds per pod on the main stem and the bottom ramification along the stem was larger than the other ramifications within one plant. Ramification‐clipping treatment increased the number of ovules per ovary (μ: 31 for control plants and 32 for clipped plants), the parameter k and the number of seeds per pod (p). This effect could be due to the competition for assimilates between the pods and seeds. Furthermore, the distribution parameters of the pollen number per stigma (m) remained stable, and the probability of pod survival (Bo) varied with different factors, including the year, pod rank and inflorescence position. Our results indicate that pollen germination is a factor that determines final seed number. This model can identify the impact of each of the factors that lead to the abortion of seeds and pods in WOSR, which include the position effect, assimilate competition and pollination limitation. However, further studies on the pollination process in WOSR should help to refine this model.     

Mapping of QTLs for morpho-agronomic and seed quality traits in a RIL population of common bean (Phaseolus vulgaris L.)

The objective of this research was to determine the quantitative trait loci (QTLs) controlling phenological traits (days to flowering, days to end of flowering, days to harvest as green pod, and days to maturity), seed size traits (seed length, seed height, seed width, and seed weight), and seed quality traits (water absorption, and coat proportion), in common bean. A population of 104 F₇ recombinant inbred lines (RILs) derived from an inter-gene pool cross between Xana, and Cornell 49242, was used to develop a genetic linkage map including 175 AFLPs, 27 microsatellites, 30 SCARs, 33 ISSRs, 12 RAPDs, 13 loci codifying for seed proteins, and the four genes Fin,fin (growth habit); Asp,asp (seed coat shininess); P,p (seed color); and I,i (resistance to bean common mosaic virus). The map has a total length of 1,042 cM distributed across 11 linkage groups aligned to those of the core linkage map of bean using common molecular markers as anchor points. The QTL analyses were carried out over three environments using the mean environment data with composite interval mapping. Thirty-one QTLs for ten traits were found to be significant in at least one environment and in the mean environment data, the number of significant QTLs identified per trait ranging from two to five. Twenty-seven of these QTLs mapped forming clusters in eight different chromosomal regions. The rationale for this clustered mapping and the possible relationship between some QTLs for phenological traits and the genes Fin and I are discussed.     

Genotypic differences in pod wall and seed growth relate to invertase activities and assimilate transport pathways in asparagus bean

Background and Aims

Coordination of sugar transport and metabolism between developing seeds and their enclosing fruit tissues is little understood. In this study the physiological mechanism is examined using two genotypes of asparagus bean (Vigna unguiculata ssp. sesquipedialis) differing in pod wall and seed growth rates. Pod growth dominates over seed growth in genotype ‘Zhijiang 121’ but not in ‘Zhijiang 282’ in which a ‘bulging pod’ phenotype is apparent from 8 d post-anthesis (dpa) onward.

Methods

Seed and pod wall growth rates and degree of pod-bulging were measured in the two genotypes together with assays of activities of sucrose-degrading enzymes and sugar content in pod wall and seed and evaluation of cellular pathways of phloem unloading in seed coat using a symplasmic fluorescent dye, 5(6)-carboxyfluorescein (CF).

Key Results

Activities of cell wall, cytoplasmic and vacuolar invertases (CWIN, CIN and VIN) were significantly smaller in pod walls of ‘282’ than in ‘121’ at 10 dpa onwards. Low INV activities were associated with weak pod wall growth of ‘282’. In seed coats, CF was confined within the vasculature in ‘282’ but moved beyond the vasculature in ‘121’, indicating apoplasmic and symplasmic phloem unloading, respectively. Higher CWIN activity in ‘282’ seed coats at 6–8 dpa correlated with high hexose concentration in embryos and enhanced early seed growth. However, CWIN activity in ‘282’ decreased significantly compared with ‘121’ from 10 dpa onwards, coinciding with earlier commencement of nuclei endoreduplication in their embryos.

Conclusions

The study shows genotypic differences between ‘bulging pod’ and ‘non-bulging’ phenotypes of asparagus bean in sucrose metabolism in relation to the pathway of phloem unloading in developing seed coats, and to pod and seed growth. Low INV activity in pod wall corresponds to its shortened and weak growth period; by contrast, the apoplasmic path in the seed coat is associated with high CWIN activity and strong early seed growth.     

Physical restriction of pods causes seed size reduction of a brassinosteroid-deficient faba bean (Vicia faba)

BACKGROUND AND AIMS: A brassinosteroid-deficient mutant faba bean (Vicia faba 'Rinrei') shows dwarfism in many organs including pods and seeds. 'Rinrei' has normal-sized seeds together with dwarf seeds, suggesting that dwarfism in the seed may be indirectly caused by brassinosteroid deficiency. The mechanism of seed size reduction in this mutant was investigated. METHODS: The associations between seed orientation in the pod, seed numbers per pod and pod lengths with seed sizes were analysed in 'Rinrei' and the wild-type plant. KEY RESULTS: 'Rinrei' seeds are tightly arranged in pods containing two or three seeds. Seed size decreased as the number of seeds per pod increased or as the length of the pod decreased. Where no physical restriction occurred between seeds in a pod, the wild-type faba bean seeds had a nearly constant size regardless of seed number per pod or pod length. 'Rinrei' seeds in pods containing single seeds were the same size as wild-type seeds. Brassinolide treatment increased the seed size and the length of pods containing three seeds in 'Rinrei'. CONCLUSION: Seed size of 'Rinrei' is mainly regulated through a reduction of pod length due to brassinosteroid deficiency; physical restriction within pods causes a reduction in seed size. These results suggest a possible mechanism for increasing faba bean yields to optimal levels.     

Rate of Increase in Size and Dry Weight of Individual Pods of Field Grown Soya Bean Plants

The dry weight of the whole fruit, the pod wall and an enclosedseed of randomly harvested soya beans is estimated from theexternal dimensions of the attached pod. The relations betweendimensions and dry weight are independent of cultivar and growthcondition and can be used on pods from 1 cm in length untilthe seeds reach their maximum fresh weight. Dimensions of tagged pods of three cultivars of field grownsoya beans differing in time to reach maturity were measuredevery 2–3 days from initial pod elongation until maturation.Dry weights for each pod were estimated from the dimensions,and the dry weight accumulation with time was fitted to thelogistic function to find the growth rate that best characterizedthe data for each pod. The final weight, the specific growthrate and the maximum growth rate of the whole fruit, the podwall and a single seed were subjected to analysis of variance. The most significant difference between pods of these cultivarswas the specific growth rate of individual seeds, which decreasedwith increasing maturity group. There were no differences ingrowth of the pod wall. However, most of the variation was betweenindividual pods within a cultivar, where the rate of dry weightaccumulation of the whole fruit, governed largely by the seedgrowth rate times the number of seeds, was highly correlatedwith the earlier growth of the pod wall. This suggests thatthe growth of individual whole fruit was determined early inpod development and was slightly influenced by factors appliedduring the period of rapid seed growth. Glycine max (L.) Merrill, Soya bean, seed growth analysis, specific growth rate     

Maternal control of seed weight in rapeseed (Brassica napus L.): the causal link between the size of pod (mother,source) and seed (offspring,sink)

Seed size/weight is one of the key traits related to plant domestication and crop improvement. In rapeseed (Brassica napus L.) germplasm, seed weight shows extensive variation, but its regulatory mechanism is poorly understood. To identify the key mechanism of seed weight regulation, a systematic comparative study was performed. Genetic, morphological and cytological evidence showed that seed weight was controlled by maternal genotype, through the regulation of seed size mainly via cell number. The physiological evidence indicated that differences in the pod length might result in differences in pod wall photosynthetic area, carbohydrates and the final seed weight. We also identified two pleiotropic major quantitative trait loci that acted indirectly on seed weight via their effects on pod length. RNA‐seq results showed that genes related to pod development and hormones were significantly differentially expressed in the pod wall; genes related to development, cell division, nutrient reservoir and ribosomal proteins were all up‐regulated in the seeds of the large‐seed pool. Finally, we proposed a potential seed weight regulatory mechanism that is specific to rapeseed and novel in plants. The results demonstrate a causal link between the size of the pod (mother, source) and the seed (offspring, sink) in rapeseed, which provides novel insight into the maternal control of seed weight and will open a new research field in plants.     

An Ontogenetic Study of Canavanine Formation in the Fruit of Jack Bean, Canavalia ensiformis (L.) DC

An ontogenetic study of canavanine formation in the fruit of jack bean, Canavalia ensiformis (L.) DC. was conducted. Evidence was presented to show that the ovary wall is the reservoir for seed canavanine. The testa possesses sufficient canavanine to account for the continued elevation in seed canavanine after the pod senesces. The seed canavanine concentration is not constant inasmuch as the canavanine content per milligram dry weight or soluble protein increases abruptly with seed growth and levels off only with the onset of fruit ripening.     

Resistance to the cowpea weevil (Callosobruchus maculatus) larva in pods of cowpea (Vigna unguiculata)

Thirty cowpea, Vigna unguiculata (L.) Walpers, cultivars were evaluated as intact pods to determine if any possessed resistance to the cowpea weevil, Callosobruchus maculatus (Fabr.). Pod resistance was measured as pre-establishment larval mortality (PreM); those larvae dying after egg hatch but before penetrating into the seeds, and as post-establishment within-seed mortality (PostM); those larvae dying after penetrating into the seeds. Among the 30 varieties examined, PreM ranged from 57.9% to 99.4% and PostM ranged from 6.7% to 82.6%. Ten varieties exhibited total intact pod mortality (mortality from egg hatch to adult emergence from the seed) greater than 95%.Physical measurements were made of several pod and seed characteristics to ascertain whether the observed pod resistance was due to seed factors, pod-wall factors, or to interactions between the pod and seeds. Although resistance to breakage (from handling plus pod dehiscence) is a necessary characteristic for overall pod protection against C. maculatus; other factors are also important. Among the other pod and seed characteristics measured to identify major resistance factors, seed coat thickness was the one most highly correlated with pod resistance. Our results suggest that interactions between pod-wall and seed coat characteristics play a large role in pod resistance of cowpeas to C. maculatus.     

Inheritance of growth habit detected by genetic linkage analysis using microsatellites in the common bean (Phaseolus vulgaris L.)

The genetic linkage map for the common bean (Phaseolus vulgaris L.) is a valuable tool for breeding programs. Breeders provide new cultivars that meet the requirements of farmers and consumers, such as seed color, seed size, maturity, and growth habit. A genetic study was conducted to examine the genetics behind certain qualitative traits. Growth habit is usually described as a recessive trait inherited by a single gene, and there is no consensus about the position of the locus. The aim of this study was to develop a new genetic linkage map using genic and genomic microsatellite markers and three morphological traits: growth habit, flower color, and pod tip shape. A mapping population consisting of 380 recombinant F10 lines was generated from IAC-UNA × CAL143. A total of 871 microsatellites were screened for polymorphisms among the parents, and a linkage map was obtained with 198 mapped microsatellites. The total map length was 1865.9 cM, and the average distance between markers was 9.4 cM. Flower color and pod tip shape were mapped and segregated at Mendelian ratios, as expected. The segregation ratio and linkage data analyses indicated that the determinacy growth habit was inherited as two independent and dominant genes, and a genetic model is proposed for this trait.     

Seed coat cell turgor responds rapidly to air humidity in chickpea and faba bean

The turgor pressure in cells of chickpea (Cicer arietinum L.) and faba bean (Vicia faba L.) seed coats was measured with a pressure probe. Measurements were made under in situ conditions by removing a section of wall from a pod, which remained attached to the plant, and exposing the intact seed. If the pod wall was removed and the turgor measurements made under ambient laboratory conditions of 50% to 70% relative humidity (RH), cell turgor pressure declined over time, typically reaching 0 MPa. If the pod wall was removed and the turgor measurements made under conditions of 100% RH, however, cell turgor pressure was stable over time, relatively uniform within the seed coat tissue, and was found to be 0.1-0.3 MPa for chickpea, and 0.1-0.2 MPa for faba bean. In both species there was a marked decline in cell turgor, beginning within about 60 s, when humidification was discontinued. The decline in cell turgor occurred regardless of the depth of the cell within the seed coat tissue, and this decline could be stopped, but not entirely reversed, when humidification was restored. An increase in cell turgor could also be caused by wetting of the seed. These responses indicate that a very rapid water exchange can occur within the seed coat tissue in situ. The rapid and, in some cases, relatively permanent loss of seed coat cell turgor in the absence of humidification raises serious concerns regarding desiccation artefacts which may be involved in the empty seed coat technique, often used to study seed carbon and water relations in grain legumes.     

Pod and Seed Mycoflora on Transgenic and Conventional Soybean [Glycine max (L.) Merrill] cultivars in Mississippi

A 2-year (1999-2000) study was conducted at Starkville and Stoneville, MS to determine if the occurrence of the mycoflora varied on Roundup Ready (transgenic) compared to conventional soybean (Glycine max) cultivars. A total of 7,658 fungal isolates were identified from the pod and seed tissues of four cultivars compared at growth stages R6 and R8. Ninety-nine percent of all fungi isolated were mitosporic fungi and ascomycetes. In both years, total fungal isolates from the two locations were greater from the pod (65%) than from seed (33%) tissues. Isolation frequency from conventional cultivars was 54% compared to 46% for the transgenic cultivars. The most common fungi identified that are reported pathogens of soybean included Alternaria, Cercospora, Cladosporium, Diaporthe, Fusarium and Verticillium spp. When main effects and interactions were compared among the frequency data for the fungal genera, significant differences occurred, but consistent trends were not noted. Isolation frequencies of Diaporthe spp. during the R6 growth stage, were significantly greater on the conventional than on the transgenic cultivars in both years of the study, but only at Starkville. Isolation frequencies from samples taken during the R8 growth stage were similar at both locations in 1999 and 2000. Fusarium spp. isolated at R6 and R8 growth stages from pod and seed tissues were significantly greater on conventional than on transgenic cultivars in 2000. Even though frequencies were often significantly different between the transgenic and conventional cultivars, the data was not consistent between locations, pod and seed tissues, or growth stages. The pod and seed mycoflora of transgenic and conventional soybean cultivars was, therefore, similar in Mississippi.     

Weedy adaptation in Setaria spp. IV. Changes in the germinative capacity of S. faberii (poaceae) embryos with development from anthesis to after abscission

Studies were conducted evaluating germinability states in giant foxtail (Setaria faberii) embryos, as well as surrounding tissues (hull, caryopsis), with germination assays. Further, seed age, fascicle arrangement, flowering patterns, and elongation in the inflorescence were evaluated. Both qualitatitive and quantitative morphological observations of the hull and the caryopsis were revealed by precisely determined fertilized spikelet age from anthesis until after seed abscission. Red coloration of the placental pad at ≈ 11 d after anthesis is probably a morphological indicator of physiological maturity. Germinability of giant foxtail embryos changed with development. Four qualitatively different types of embryo germination were observed during development of the seed: early disorganized callus growth at the basal, coleorhizal end of the embryo; germination of immature embryos with shortened and thickened axes; germination of the scutellum; and germination and growth of the coleoptile and coleorhiza in embryos aged 7 d after anthesis and older. Axis-specific embryo germinability was also observed. Inhibition of the embryo could be localized to the coleoptile, the coleorhiza, or both. These studies provide evidence for a complex model of germinability regulation based on the independent, asynchronous actions of the embryo, caryopsis, and hull compartments, as well as on their dependent, synchronous action. These studies provide evidence for a dynamic, developmental model of giant foxtail germinability regulation resulting in phenotypes with a wide range of germinability shed from an individual panicle. These diverse germinability phenotypes are found at all stages of development, but particularly when the seed is shed and the soil seed bank is replenished.     

Genetic variation underlying pod size and color traits of common bean depends on quantitative trait loci with epistatic effects

Common bean is an important vegetable legume in many regions of the world. Size and color of fresh pods are the key factors for deciding the commercial acceptance of bean as a fresh vegetable. The genetic basis of important horticultural traits of common bean is still poorly understood, which hinders DNA marker-assisted breeding in this crop. Here we report the identification of single-locus and epistatic quantitative trait loci (QTLs), as well as their environment interaction effects for six pod traits, namely width, thickness, length, size index, beak length and color, using an Andean intra-gene pool recombinant inbred line population from a cross between a cultivated common bean and an exotic nuña bean. The QTL analyses performed detected a total of 23 QTLs (single-locus QTLs and epistatic QTLs): five with only individual additive effects and six with only epistatic effects, while the remaining twelve showed both effects. These QTLs were distributed across linkage groups (LGs) 1, 2, 4, 6, 7, 8, 9, 10 and 11; particularly noteworthy are the QTLs for pod size co-located on LGs 1 and 4, indicative of tight linkage or genes with pleiotropic effects governing these traits. Overall, the results obtained showed that additive and epistatic effects are the major genetic basis of pod size and color traits. The mapping of QTLs including epistatic loci for the six pod traits evaluated provides support for implementing marker-assisted selection toward genetic improvement of common bean.     

Regulation of Leaflet Photosynthesis by Developing Fruit in the Pea

Changes in the net photosynthesis of leaflets of Pisum sativum L. (cv. Onward) were found to reflect closely the pattern of assimilate demand in subtended fruit during development. An early peak in leaflet photosynthesis corresponded with the rapid elongation of the fruit; a second peak with the inflation of the pod and initiation of seed growth. However, where the subtended fruit was removed at anthesis, the pattern of fluctuation in leaflet photosynthesis was altered, while leaflets at vegetative nodes maintained a lower photosynthetic rate which fluctuated little. The pattern of net photosynthesis in the fruit appears to reflect the high respiration rates associated with pod elongation and seed growth. This closely coupled system, where fruit assimilate demand modulates the photosynthesis of adjacent leaflets, provides a scope for study of source-sink relationships with the minimum of experimental manipulation.     

Assessment of chemical resistance of wild and cultivated Vigna species to the brown pod-bug Clavigralla tomentosicollis Stal (Hem., Coreidae)

The effects of secondary metabolites in different Vigna species on the development of Clavigralla tomentosicollis were investigated in an artificial seed system using different fractions of crude pod extracts, while the orientation response of this pod‐bug to volatile extracts was studied using a dual‐choice olfactometer. Feeding on the neutral fraction extracts, in contrast to the basic and acidic fractions, resulted in significantly higher mortalities, longer total developmental time, and lower growth index of the insects in comparison with controls. All volatile extracts elicited an avoidance reaction by C. tomentosicollis, except the volatile from the susceptible genotype IT84S‐2246 which generally attracted as many insects as controls. Extracts from wild Vigna species showed higher activity than those from their cultivated relatives. The present study which has established that most secondary metabolites in cowpea pods were localized in the neutral fraction of the crude extract, could facilitate experiments on the separation and characterization of the toxic factors involved.     

Effect and interaction of temperature and photoperiod on growth and partitioning in three groundnut (Arachis hypogaea L.) genotypes1

Effect of temperature and photoperiod and their interaction on plant growth and partitioning of dry matter to pods was examined in three selected groundnut genotypes viz., TMV 2, NC Ac 17090 and VA 81B. The genotypes were grown in six walk-in growth chambers which were programmed to simulate three temperature regimes (22/18°C, 26/22°C and 30/26°C day/night) each under long (12 h) and short (9 h) photoperiods. The plant growth rates and partitioning of dry matter to pods were estimated on a thermal time basis. Plant growth rate (PLGR) was significantly influenced by temperature, photoperiod and genotype, whereas pod growth rate (PDGR) was influenced primarily by temperature and genotype. The interaction of genotype with photoperiod and with temperature was significant for both PLGR and PDGR. For example, at the 22/18°C temperature regime, VA 81B had a high PDGR, while NC Ac 17090 did not even initiate pod growth. The partitioning of dry matter to pods (Pf) was also significantly influenced by photoperiod, temperature and genotype, and significant interactions were found. Photoperiod did not significantly affect Pf under the low temperature regime, but at higher temperatures, partitioning to pods was significantly greater under short days. Pf of VA 81B was relatively insensitive to photoperiod compared with the other two genotypes. The study provided evidence of genotypic variability for photoperiod × temperature interactions which could influence adaptation of groundnut genotypes to new environments.