D After Anthesis Research Articles

Awn contribution to gas exchanges of barley ears

The effects of awn removal on ear gas exchange in four barley lines (Morex, Harrington, Steptoe, and TR306) were studied under a controlled environment using a Before-After Control-Impact Paired (BACIP) experimental design. From ear emergence to grain maturity, plants were grown in pots at either 60 or 90 % of soil water holding capacity. Gas-exchange measurements of ears were made 9 and 10 d after anthesis (DAA). On 11 DAA, awn removal was performed on half of the ears in each pot, followed by measurements on both intact and de-awned ears on 12 and 13 DAA. Net photosynthetic (P N) and transpiration (E) rates decreased significantly with awn removal, but dark respiration (R D) rate was not affected. We estimated for each ear a temperature-adjusted respiration rate (R a) from R D. When we corrected P N with R a, we found that rates of spikelet photosynthesis were largely underestimated. Moderate water stress had minimal effect on gas exchange of bracts and awns of the barley ear. Barley lines did not differ for any individual gas-exchange parameter.

Variations in sugars and cyclitols during development and maturation of seeds of brazilwood (Caesalpinia echinata Lam., Leguminosae)

Seeds of Caesalpinia echinata are tolerant to desiccation, keeping their viability up to 0.08 g H2O g-1 DM. When stored under laboratory conditions they lose their viability within one month, while under low temperatures germination ability is maintained during two years of storage. Here, we report on the variations of soluble carbohydrates, cyclitols and cyclitol galactosides in seeds of C. echinata during development and maturation, aiming for a better understanding of the involvement of these compounds in the maturation process and seed quality. Individual flowers were tagged on the day of anthesis and pods were collected directly from the branches from 32 to 59 d after anthesis (DAA). Seeds were also collected after shedding. The physiological maturity of C. echinata seeds occurred 60-65 DAA, immediately before shedding, when seeds had 0.43-0.67 g H2O g-1 DM. Soluble carbohydrates made up about 10% of the seed dry mass. Gas chromatography and mass spectrometry allowed the identification and quantification of 12 soluble carbon compounds present in both the axis and cotyledons of developing C. echinata seeds. These included fructose, glucose, sucrose, raffinose, the cyclitols myo-inositol, D-chiro-inositol, D-pinitol, and the cyclitol galactosides, galacto-pinitol A, galacto-pinitol B and ciceritol. Sucrose was present in high proportions in both tissues during seed development. Traces of raffinose were also detected, mainly in immature seeds. D-chiro-inositol and myo-inositol were found in low amounts and pinitol transiently increased in both axis and cotyledons. Ciceritol, galacto-pinitol A and galacto-pinitol B accumulated from 40 DAA to the end of seed maturation, representing along with sucrose the major carbon reserves of C. echinata seeds. These results allowed us to conclude that variations in soluble carbon reserves of C. echinata seeds were related to the maturation process and seed quality.

Gene expression of stearoyl‐ACP desaturase and Δ12 fatty acid desaturase 2 is modulated during seed development of flax (Linum usitatissimum)

Flax's recent popularity in human and animal foods is mostly due to its desirable FA composition. Flax is an excellent source of omega-3 FA, which have been shown to have many health benefits. To date, little is known about the genetic and environmental factors that control the FA composition of flax seeds. To elucidate some of the important genetic components, reverse transcriptase (RT)-PCR and real-time PCR were used to determine the expression profiles of two key FA biosynthetic genes during seed development. Plants of flax cultivar AC McDuff were grown under field conditions, and RNA was extracted from ovaries and developing bolls collected from 2 d after anthesis (DAA) to maturity. Desaturation enzymes stearoyl-ACP desaturase (SAD) and delta12 FA desaturase 2 (FAD2) were both expressed in ovaries, and their expression was differentially modulated throughout seed development. SAD was most highly expressed in ovaries. Its expression quickly decreased until 4 DAA; this was followed by a slight peak at 8 DAA, only to return to relatively low levels of expression in maturing bolls, ranging from 2.1% to 4.5% relative to the level observed in ovaries. FAD2 expression displayed a different temporal pattern. While expression of FAD2 did decrease in the early stages of seed development, expression increased starting at 8 DAA, peaking at 16 DAA, when it was 158% relative to the level observed in ovaries. FAD2, which desaturates oleic acid (18:1cisdelta9) into linoleic acid (18:2cisdelta9,12), is therefore controlled at the transcription level. To relate enzyme expression with FA profile, GC was performed on the same subsamples used for RT-PCR and real-time PCR, and proportions of palmitic, stearic, oleic, linoleic, and linolenic acids were determined for the same developmental stages. Although FAD2 expression increased from 8 to 16 DAA, relative changes in linoleic acid (18:2cis delta9,12) were not observed. However, linolenic acid (ALA; alpha-18:3; 18:3cisdelta9,12,15) levels increased steadily, meaning that linoleic acid (18:2cisdelta9,12) is a transient substrate converted by FAD3 as quickly as it is produced by FAD2. Phenotypes are the result of genotypes, environment, and the interaction of the two. To evaluate the environmental impact on the production of FA in flax, FA profiles were assessed in a total of four environments (two locations, two years). Warm and dry environmental conditions resulted in lower levels of PUFA 18:2cisdelta9,12 and 18:3cisdelta9,12,15, and higher levels of 18:1 cisdelta9. FAD2 expression and/or activity may therefore be affected by the environment.

Cell wall modifications in the pericarp of litchi (Litchi chinensis Sonn.) cultivars that differ in their resistance to cracking

SummaryChanges in structural calcium and galacturonan concentrations, the degree of methylesterification of pectins, the activities of pectin methylesterase (PME) and soluble and wall-bound peroxidases (POD), and amino acid compositions of structural proteins in the fruit pericarp (skin) were studied in cracking-resistant ‘Huaizhi’ and cracking-susceptible ‘Nuomici’ litchi. Structural calcium concentrations were higher in ‘Huaizhi’ than in ‘Nuomici’ and decreased from 22 d to 52 d after anthesis (DAA), then increased. Galacturonan concentrations increased over time and were higher in ‘Huaizhi’. Methylesterification of pectins increased from 50% at 22 DAA, to 55% at 78 DAA in ‘Huaizhi’, but fluctuated around 48% in ‘Nuomici’. PME activity was significantly higher in ‘Nuomici’ than in ‘Huaizhi’ in the later stages of fruit development (> 50 DAA). The activities of soluble (SPOD) and ionically wall-bound POD (IWBPOD) increased with fruit development. Maximum SPOD activity was similar in both cultivars, whereas the peak activity of IWBPOD was higher in ‘Nuomici’ than in ‘Huaizhi’. Total concentrations of SDS-soluble amino acids were similar in the two cultivars. Total structural proteins (SDS-insoluble amino acids) decreased with fruit development. Among the SDS-insoluble amino acids, hydroxyproline was the only one that increased over time. The results suggest that higher levels of structural calcium and galacturonans may contribute to cracking resistance in ‘Huaizhi’, while a higher activity of IWBPOD, which catalyses the formation of phenolic cross-linkages and an irreversible increase in the rigidity of cell walls, may be associated with cracking susceptibility in ‘Nuomici’. Structural proteins may not be involved in resistance to cracking in litchi cultivars.

Girdling increases carbohydrate availability and fruit-set in citrus cultivars irrespective of parthenocarpic ability

SummaryThe effects of girdling performed at various dates were evaluated during two consecutive years in high- and low-bearing commercial orchards of ‘Fortune’ mandarin and ‘Clausellina’ Satsuma mandarin. The time-dependent response was evaluated through fruitlet abscission, final fruit-set and yield as related to carbohydrate contents in developing fruitlets. A few days after treatment, girdling increased the soluble sugars content (SSC) in fruitlets, reduced the daily fruit drop, and thereby diminished abscission. Application of girdling to low-bearing ‘Fortune’ mandarin orchards was most effective 15 d before anthesis (DBA) and 35 d after anthesis (DAA). It increased yield by 125%. In high-bearing orchards, the best results were achieved by girdling 35 DAA, which increased yield by 28%. In both orchards, the effect of girdling declined gradually from 35 DAA to the end of fruit drop. SSC values were found to be positively correlated with fruit-set at 25 DAA, but this tendency was reversed by 50 DAA. This study showed that the effects of girdling are time-dependent, and that yield can be improved in cultivars of different parthenocarpic ability.

The presence of oxalate in the pericarp and fruit pedicel is not linked to a shortage of fruit calcium and increase in cracking incidence in litchi

SummaryConcentrations of oxalate and calcium in soluble or structural forms, as calcium oxalate in the pericarp, or as oxalate in the fruit pedicel, and the distribution of calcium in the pericarp and fruit pedicel were studied in litchi (Litchi chinensis Sonn.) using cracking-resistant cv. ‘Huaizhi’ and cracking-susceptible cv. ‘Nuomici’. Oxalate in the pericarp existed mainly in a soluble form and was significantly (P < 0.05) higher in ‘Huaizhi’ than in ‘Nuomici’. The concentration of oxalate was higher than the amount of calcium oxalate. Calcium was first mobilised for the construction of cell walls in the pericarp, despite the high concentration of oxalate in the tissue. ‘Huaizhi’ fruit accumulated more structural calcium and calcium oxalate in the pericarp than ‘Nuomici’ fruit. There was no difference in soluble calcium between the two cultivars. Calcium oxalate was located chiefly in the epidermis of the pericarp, and increased in concentration within 50 d after anthesis (DAA) in ‘Nuomici’ and 64 DAA in ‘Huaizhi’, then decreased as structural calcium increased, indicating that calcium sequestered by oxalate could be remobilised.The fruit pedicel had higher concentrations of oxalate than the pericarp. X-ray microanalysis showed abundant calcium in pedicel tissue adjoining the fruit base, and less calcium in tissues at the fruit base and in pedicel tissue 1 cm from the fruit, suggesting that the pedicel serves as a bottleneck to calcium movement. This bottleneck is not likely to be caused by oxalate in the pedicel. Oxalate in the pericarp and in the fruit pedicel is not linked to a shortage of fruit calcium or to an increase in cracking incidence in litchi.

Sugar metabolism in developing lupin seeds is affected by a short-term water deficit

A short-term water deficit (WD) imposed during the pre-storage phase of lupin seed development [15-22 d after anthesis (DAA)] accelerated seed maturation and led to smaller and lighter seeds. During seed development, neutral invertase (EC 3.2.1.26) and sucrose synthase (EC 2.4.1.13) have a central role in carbohydrate metabolism. Neutral invertase is predominant during early seed development (up to 40 DAA) and sucrose synthase during the growing and storage phase (40-70 DAA). The contribution of acid invertase is marginal. WD decreased sucrose synthase activity by 2-fold and neutral invertase activity by 5-6-fold. These changes were linked to a large decrease in sucrose ( approximately 60%) and an increase of the hexose:sucrose ratio. Rewatering restored sucrose synthase activity to control levels while neutral invertase activity remained depressed (30-60%). A transient accumulation of starch observed in control seeds was abolished by WD. Despite the several metabolic changes the final seed composition was largely unaltered by WD except for approximately 60% increase in stachyose and raffinose (raffinose family oligosaccharides). This increase in raffinose family oligosaccharides appears as the WD imprinting on mature seeds.

Comparison of changes in fruit gene expression in tomato introgression lines provides evidence of genome-wide transcriptional changes and reveals links to mapped QTLs and described traits

Total soluble solids content is a key determinant of tomato fruit quality for processing. Several tomato lines carrying defined introgressions from S. pennellii in a S. lycopersicum background produce fruit with elevated Brix, a refractive index measure of soluble solids. The genetic basis for this trait can be determined by fine-mapping each QTL to a single gene, but this is time-consuming and technically demanding. As an alternative, high-throughput analytical technologies can be used to provide useful information that helps characterize molecular changes in the introgression lines. This paper presents a study of transcriptomic changes in six introgression lines with increased fruit Brix. Each line also showed altered patterns of fruit carbohydrate accumulation. Transcriptomic changes in fruit at 20 d after anthesis (DAA) were assessed using a 12 000-element EST microarray and significant changes analysed by SAM (significance analysis of microarrays). Each non-overlapping introgression resulted in a unique set of transcriptomic changes with 78% of significant changes being unique to a single line. Principal components analysis allowed a clear separation of the six lines, but also revealed evidence of common changes; lines with quantitatively similar increases in Brix clustered together. A detailed examination of genes encoding enzymes of primary carbon metabolism demonstrated that few of the known introgressed alleles were altered in expression at the 20 DAA time point. However, the expression of other metabolic genes did change. Particularly striking was the co-ordinated up-regulation of enzymes of sucrose mobilization and respiration that occurred only in the two lines with the highest Brix increase. These common downstream changes suggest a similar mechanism is responsible for large Brix increases.

Pod set related to photosynthetic rate and endogenous ABA in soybeans subjected to different water regimes and exogenous ABA and BA at early reproductive stages.

The physiological reasons for reduced pod set in soybean (Glycine max) caused by drought during anthesis are not established. The objective of this study was to investigate the involvement of photosynthetic rate (A), pod endogenous abscisic acid (ABA) and exogenously applied ABA and 6-benzylaminopurine (BA) in regulating pod set in soybean during drought. Two pot experiments were done in a controlled-environment glasshouse. In expt I, soybeans were either well-watered (WW) or droughted by withholding water from 4 d before to 4 d after anthesis (DAA). In expt II, soybeans were drought-stressed (DS) from -4 to 4 DAA. From -2 to 4 DAA, some of the WW and DS plants were treated with 0.1 mm ABA or 1 mm BA. Drought stress decreased A, but increased pod ABA concentration ([ABA]). Pod set decreased only when A had decreased by 40 %, and pod [ABA] had increased 1.5-fold. Beyond the thresholds, pod set correlated positively with A and negatively with pod [ABA]. Exogenously applied ABA decreased A and pod set in WW plants, whilst it increased A and pod set in DS plants; exogenous BA had opposite effects. In these plants, pod set correlated linearly with A. Drought-induced decrease in A is significant in inducing pod abortion, probably as a consequence of carbohydrate deprivation. The effects of ABA and BA on pod set may be partially due to their effects on photosynthate supply.

Fusarium graminearum Infection during Wheat Seed Development and Its Effect on Seed Quality

Fusarium head blight (FHB) caused by Fusarium graminearum Schwabe causes losses in wheat (Triticum aestivum L.) seed quality and yield. Field studies were conducted in 2000 and 2001 to investigate F. graminearum seed infection (SI) and its relationship to seed germination and vigor and the production of the mycotoxin deoxynivalenol (DON). Seeds from four soft red winter wheat cultivars having different levels of Type II resistance to F. graminearum were harvested at frequent intervals during development and maturity. Low levels of SI occurred late in seed development in 2000, which resulted in high seed quality. In 2001, under heavy disease pressure, F. graminearum increased from &lt;20% at 10 d after anthesis (DAA) to &gt;98% at maturity in both resistant and susceptible cultivars, resulting in unacceptable standard germination (SG) (&lt;80%) and seed vigor [&lt;70% accelerated aging (AA) germination] early in seed development. Although fungicide seed treatment improved the SG of all cultivars, it still remained below acceptable commercial seed quality (80%). Deoxynivalenol was present at significant levels (5 to 25 mg kg−1) throughout seed development and maturation in all cultivars in 2001. Although the resistant cultivar P25R18 had reduced levels of DON and lower seed damage compared with susceptible P2552, resistance had no effect on SI and seed quality. Therefore, under severe disease pressure, Type II resistance offers little advantage to seed producers in reducing SI and improving seed quality during severe FHB epidemics.

Dry matter accumulation in citrus fruit is not limited by transport capacity of the pedicel.

The vascularization of the pedicel in Marisol clementine (Citrus clementina Hort. ex Tanaka) has been characterized in relation to fruit growth. Phloem and xylem formation occurred during the first half of the period of fruit growth. Phloem cross-sectional area reached its maximum value by the end of fruitlet abscission, 78 d after anthesis (DAA), shortly after the rate of accumulation of dry matter in fruitlets reached its maximum value. Secondary xylem formation occurred until day 93, well after the end of fruitlet abscission. At fruit maturity, xylem accounted for 42-46 % of the cross-section of the pedicel. Vessels differentiated in this late-formed xylem. Formation of phloem and early xylem was directly related to fruitlet size (and growth rate). Differences in the rate of formation of conductive tissues in the pedicel of the developing fruitlets followed rather than preceded the differences in growth rate. Specific mass transfer (SMT) in the phloem was highest in the fastest growing fruitlets, and peaked during the late stages of fruitlet abscission (72-78 DAA) and during the main period of fruit growth (107-121 DAA). Application of a synthetic auxin to developing fruits, either at the end of flowering (2,4-D) or by day 64 after flowering (2,4-DP), increased the growth rate of the fruit and fruit size at maturity (8-13 % increase in fruit diameter at maturity). These auxin applications also enhanced the formation of conductive tissues in the pedicel, with a specific effect on phloem formation. Applying auxin at flowering resulted in a reduction in the phloem SMT by days 72-78, whereas auxin application on day 64 increased this parameter. Despite this difference in behaviour, which resulted from the different time-course of the growth response of the fruit to auxin applications, these applications increased fruit size to a similar extent. Severing 37 % of the phloem of the pedicel during the main period of fruit growth resulted in an increase in the specific mass transfer in the phloem but had no influence on fruit growth. These observations demonstrate that the transport capacity in the phloem of the pedicel does not limit fruit growth and, within the limits of our experiments, an increase in demand by the fruit appeared to be matched by an increase in SMT. The dependence of late xylem formation (after the period of fruitlet abscission) on fruitlet growth was demonstrated in Salustiana orange [Citrus sinensis (L.) Osbeck] by means of controlling fruit growth through the manipulation of leaf area. Fruit growth at this time was more closely related to leaf area than to carbohydrate levels, suggesting that it may be limited by current photosynthesis.

Hormone and seed-specific regulation of pea fruit growth.

Growth of young pea (Pisum sativum) fruit (pericarp) requires developing seeds or, in the absence of seeds, treatment with gibberellin (GA) or auxin (4-chloroindole-3-acetic acid). This study examined the role of seeds and hormones in the regulation of cell division and elongation in early pea fruit development. Profiling histone H2A and gamma-tonoplast intrinsic protein (TIP) gene expression during early fruit development identified the relative contributions of cell division and elongation to fruit growth, whereas histological studies identified specific zones of cell division and elongation in exocarp, mesocarp, and endocarp tissues. Molecular and histological studies showed that maximal cell division was from -2 to 2 d after anthesis (DAA) and elongation from 2 to 5 DAA in pea pericarp. Maximal increase in pericarp gamma-TIP message level preceded the maximal rate of fruit growth and, in general, gamma-TIP mRNA level was useful as a qualitative marker for expanding tissue, but not as a quantitative marker for cell expansion. Seed removal resulted in rapid decreases in pericarp growth and in gamma-TIP and histone H2A message levels. In general, GA and 4-chloroindole-3-acetic acid maintained these processes in deseeded pericarp similarly to pericarps with seeds, and both hormones were required to obtain mesocarp cell sizes equivalent to intact fruit. However, GA treatment to deseeded pericarps resulted in elevated levels of gamma-TIP mRNA (6 and 7 DAA) when pericarp growth and cell enlargement were minimal. Our data support the theory that cell division and elongation are developmentally regulated during early pea fruit growth and are maintained by the hormonal interaction of GA and auxin.

Activities of starch hydrolytic enzymes and sucrose-phosphate synthase in the stems of rice subjected to water stress during grain filling.

To understand the effect of water stress on the remobilization of prestored carbon reserves, the changes in the activities of starch hydrolytic enzymes and sucrose-phosphate synthase (SPS) in the stems of rice (Oryza sativa L.) during grain filling were investigated. Two rice cultivars, showing high lodging-resistance and slow remobilization, were grown in the field and subjected to well-watered (WW, psi(soil)=0) and water-stressed (WS, psi(soil)=-0.05 MPa) treatments 9 d after anthesis (DAA) till maturity. Leaf water potentials of both cultivars markedly decreased during the day as a result of WS treatment, but completely recovered by early morning. WS treatment accelerated the reduction of starch in the stems, promoted the reallocation of prefixed (14)C from the stems to grains, shortened the grain filling period, and increased the grain filling rate. More soluble sugars including sucrose were accumulated in the stems under WS than under WW treatments. Both alpha- and beta-amylase activities were enhanced by the WS, with the former enhanced more than the latter, and were significantly correlated with the concentrations of soluble sugars in the stems. The other two possible starch-breaking enzymes, alpha-glucosidase and starch phosphorylase, showed no significant differences in the activities between the WW and WS treatments. Water stress also increased the SPS activity that is responsible for sucrose production. Both V(limit) and V(max), the activities of the enzyme at limiting and saturating substrate concentrations, were enhanced and the activation state (V(limit)/V(max)) was also increased as a result of the more significant enhancement of V(limit). The enhanced SPS activity was closely correlated with an increase of sucrose accumulation in the stems. The results suggest that the fast hydrolysis of starch and increased carbon remobilization were attributed to the enhanced alpha-amylase activity and the high activation state of SPS when the rice was subjected to water stress.

Fate of photosynthates from spur leaves of ‘Nijisseiki’ pear during the period of rapid fruitgrowth

SummarySpurs of ‘Nijisseiki’ pear (Pyrus pyriflora Nakai) were allowed to assimilate 10033;CO2 at 87.d after anthesis (DAA) and at 108 DAA during the period of rapid fruit growth. Then the spurs were sampled periodically until fruit harvest time to trace the time course and amount of movement of assimilates from spur leaves to individual organs in the spur. The amount of 10033;C absorbed by fruit within 3.d after labelling was constant until harvest, regardless of the labelling date. However, the total amount of 10033;C in the spur decreased continually until harvest.Of the total amount of 10033;C recovered in the spur labelled at 87 DAA, by harvest, 43.2% of 13C was found in the fruit flesh (cortex of receptacle), 5% in the core (pith of receptacle 1 pericarp 1 seeds), 5.6% in spur stem, 5.4% in the source leaves, and 40.8% was respired and exported from the spur. Of total amount of initial 10033;C labelled at 108 DAA, at harvest, the proportion of 10033;C translocated to flesh, core, stem, respired, and remaining in leaves was 61%, 6.1%, 3.0%, 24% and 5.4%, respectively. Photosynthates fixed by the spur early in the rapid growth stage of the fruit contribute more to the formation of starch and structural materials and less to solublecarbohydrates in fruit than do those fixed later.

Changes of endogenous hormone level in pollinated and N-(2-chloropyridyl)-N 9-phenylurea (CPPU)-induced parthenocarpicfruits of Lagenaria leucantha

SummaryPollinated and CPPU (N-(2-chloro-4-pyridyl)-N 9-phenylurea)-induced parthenocarpic fruits were used to study their roles in fruit set, and the metabolism of endogenous hormones in fruit of Lagenaria leucantha. Unpollinated fruit failed to develop and eventually aborted. CPPU induced parthenocarpic fruit significantly larger than pollinated ones. IAA (indoleacetic acid) and t-zeatin levels in pollinated fruit increased with fruit growth and reached their maximum values 9.d after anthesis (DAA). IAA and t-zeatin levels in CPPU-induced parthenocarpic fruit, however, declined gradually within fruit growth and were much lower than in pollinated fruit. These results suggest that CPPU did not induce parthenocarpic fruit growth by increasing the IAA level in fruit, but the effect was direct.

Influence of high temperature during pre- and post-anthesis stages of floral development on fruit-set and pollen germination in peanut

Peanut (Arachis hypogaea L.) crops are often exposed to day temperatures &gt; 35°C for short periods duringflowering, resulting in lower yields. Research was conducted to study and quantify the effects of short episodes (1– 6 d) of high temperatures during the pre- and post-anthesis stages of floral development on fruit-set, pollen viability, germination and tube growth. Plants of peanut cv. ICGV-86015 were grown in controlled environments at 28/22°C (day/night). High daytime air temperature treatments ranging from 28 (control) to 48°C were imposed at different times between 6 d before anthesis (DBA) and 6 d after anthesis (DAA) for 1, 3 or 6 d. Floral buds or flowers were tagged at different stages to determine fruit-set. Exposure to bud (tissue) temperature ≥ 39°C for 1 d significantly reduced fruit-set compared to the control at 28°C, and the magnitude of the reduction varied with stage of floral development. Floral buds were most sensitive to high temperature at 4 DBA and at anthesis, coinciding with micro-sporogenesis and pollination or fertilisation, respectively. The critical bud temperature at these stages was 33°C, above which fruit-set was reduced by 6% °C –1 . Lower fruit-set due to high temperatures at pre-anthesis and anthesis stages were due to pollen sterility and retarded pollen tube growth, respectively.

The role of leaves in citrus fruitlet abscission: Effects on endogenous gibberellin levels and carbohydrate content

SummaryIn satsuma (Citrus unshiu, cv. Clausellina) mandarin trees, defoliation shortly after anthesis of old (previous season) or young (current season) leaves increased fruitlet abscission (up to 60%) during the following 12 weeks.Defoliation also advanced the beginning of abscission. In foliated plants, maximum rates of abscission were reached 41 d after anthesis (DAA), whereas in plants without old or young leaves maximum abscission rates occurred 32 and 54 DAA, respectively. Gibberellin (GA) and carbohydrate concentrations were determined by GC-MS and HPLC, respectively. Defoliation didnot substantially modify the endogenous levels of the active GA1, although the precursors and products of this GA changed mostly at 32 DAA. Thereafter, GA concentrations were similar in foliated and defoliated plants. Carbohydrate concentrations were low, and the highest differences were observed at 21 DAA. At this time, sucrose and hexoses were slightly lower and higher, respectively, in fruitlets on defoliated plants. Higher abscission rates of fruitlet of leafless inflorescences were also associated with reduced amounts of sucrose and higher hexose concentrations during this period.The data suggest that removal of either old or young leaves advanced and increased abscission, and reduced the sucrose/hexose ratio of fruitlets during the initial period of fruit set.

Improvement of Direct Somatic Embryogenesis in Rice by Selecting the Optimal Developmental Stage of Explant and Applying Desiccation Treatment

SummaryBy selecting the optimal developmental stage of zygotic embryos used as expiants and applying desiccation treatment, we improved direct somatic embryogenesis in rice scutellum from two cultivars, Nipponbare and Sasanishiki. Zygotic embryos isolated 14-17, 21, 28-30 and 35-40 d after anthesis (DAA) from Nipponbare and 14-17, 18-21, 28-30 and 40-42 DAA from Sasanishiki were cultured on the embryo induction medium (EIM). Then they were transferred to embryo maturation medium (EMM) and germinated on the embryo germination medium (EGM). Only immature zygotic embryo isolated 14-17 DAA from Nipponbare and Sasanishiki could develop somatic embryos that germinated. Explants from embryos at other developmental stages could develop somatic embryos only until the elongating or scutellar stage. They enlarged and formed callus without further development. The EIM and EMM consisted of N6 macronutrients, B5 micronutrients, and B5 vitamins, supplemented with 0.1 g L-1 casein hydrolysate, 1.5 g L-1 proline and 1 g L-1 MES buffer. EGM consisted of MS macro- and micronutrients and MS vitamins without organic supplement. In addition, 2 mg L-1 2,4-D was added to EIM, 1 mg L-1 2,4-D to EMM and 0.01 mg L-1 zeatin to EGM. Developmental processes of somatic embryos derived from the expiants were observed by scanning electron microscopy. Desiccation treatment of maturing somatic embryo was proved to produce fully mature somatic embryos capable of germinating vigorously.

Temporal and spatial patterns of accumulation of the transcript of Myo-inositol-1-phosphate synthase and phytin-containing particles during seed development in rice.

Myo-inositol-1-phosphate (I[1]P) synthase (EC 5.5.1.4) catalyzes the reaction from glucose 6-phosphate to I(1)P, the first step of myo-inositol biosynthesis. Among the metabolites of I(1)P is inositol hexakisphosphate, which forms a mixed salt called phytin or phytate, a storage form of phosphate and cations in seeds. We have isolated a rice (Oryza sativa L.) cDNA clone, pRINO1, that is highly homologous to the I(1)P synthase from yeast and plants. Northern analysis of total RNA showed that the transcript accumulated to high levels in embryos but was undetectable in shoots, roots, and flowers. In situ hybridization of developing seeds showed that the transcript first appeared in the apical region of globular-stage embryos 2 d after anthesis (DAA). Strong signals were detected in the scutellum and aleurone layer after 4 DAA. The level of the transcript in these cells increased until 7 DAA, after which time it gradually decreased. Phytin-containing particles called globoids appeared 4 DAA in the scutellum and aleurone layer, coinciding with the localization of the RINO1 transcript. The temporal and spatial patterns of accumulation of the RINO1 transcript and globoids suggest that I(1)P synthase directs phytin biosynthesis in rice seeds.

Callose deposition is responsible for apoplastic semipermeability of the endosperm envelope of muskmelon seeds

Semipermeable cell walls or apoplastic "membranes" have been hypothesized to be present in various plant tissues. Although often associated with suberized or lignified walls, the wall component that confers osmotic semipermeability is not known. In muskmelon (Cucumis melo L.) seeds, a thin, membranous endosperm completely encloses the embryo, creating a semipermeable apoplastic envelope. When dead muskmelon seeds are allowed to imbibe, solutes leaking from the embryo are retained within the envelope, resulting in osmotic water uptake and swelling called osmotic distention (OD). The endosperm envelope of muskmelon seeds stained with aniline blue, which is specific for callose (beta-1,3-glucan). Outside of the aniline-blue-stained layer was a Sudan III- and IV-staining (lipid-containing) layer. In young developing seeds 25 d after anthesis (DAA) that did not exhibit OD, the lipid layer was already present but callose had not been deposited. At 35 DAA, callose was detected as distinct vesicles or globules in the endosperm envelope. A thick callose layer was evident at 40 DAA, coinciding with development of the capacity for OD. Removal of the outer lipid layer by brief chloroform treatment resulted in more rapid water uptake by both viable and nonviable (boiled) seeds, but did not affect semipermeability of the endosperm envelope. The aniline-blue-staining layer was digested by beta-1,3-glucanase, and these envelopes lost OD. Thus, apoplastic semipermeability of the muskmelon endosperm envelope is dependent on the deposition of a thick callose-containing layer outside of the endosperm cell walls.