Co-ordinate regulation of sterol biosynthesis enzyme activity during accumulation of sterols in developing rape and tobacco seed

The activities of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, sterol methyl transferase 1 and sterol acyltransferase, key enzymes involved in phytosterol biosynthesis were shown to be co-ordinately regulated during oilseed rape ( Brassica napus L.) and tobacco ( Nicotiana tabacum L.) seed development. In both plants, enzyme activities were low during the initial stages of seed development, increasing towards mid-maturation where they remained stable for a time, before declining rapidly as the oilseeds reached maturity. During seed development, the level of total sterols increased 12-fold in tobacco and 9-fold in rape, primarily due to an increase in steryl ester production. In both seed tissues, stages of maximum enzyme activity coincided with periods of high rates of sterol production, indicating developmental regulation of the enzymes to be responsible for the increases in the sterol content observed during seed development. Consistent with previous studies the data presented suggest that sterol biosynthesis is regulated by two key steps, although there may be others. The first is the regulation of carbon flux into the isoprenoid pathway to cycloartenol. The second is the flux from cycloartenol to Delta(5)-end-product sterols. The implications of the results in terms of enhancing seed sterol levels by genetic modification are also discussed.     

Development of pepper (Capsicum annuum) seed quality

Changes in seed quality in pepper (Capsicum annuum L.) were monitored during seed development and maturation in two seasons. Seed quality was assessed by a number of different tests, but principally by determining seed storage longevity in laboratory tests and seedling growth in glasshouse tests. Mass maturity (defined as the end of the seed-filling phase) occurred 49–53 days after anthesis (DAA) in 1989 (varying among fruit layers) and 53 DAA in 1990 when seed moisture contents were 51–53%. The onset of both germinability and desiccation tolerance occurred either just before or at mass maturity. Maximum potential longevity (assessed by the value of the seed lot constant K_i) was achieved 63–65 DAA, i.e. not until 10–12 days after mass maturity (DAMM), in both years. Seedling dry weights in the glasshouse growth tests were maximal later still - for seeds harvested 17–21 DAMM in 1989 and 17 DAMM in 1990; the effects on seedling weight arose from differences in times from sowing to emergence (P < 0.005) among different seed harvests, with no significant differences in subsequent relative growth rates (P > 0.25). Seed priming reduced mean germination times for seeds harvested at all stages of development, but had little effect on germination capacity and potential longevity, and did not affect the pattern of changes in potential longevity during seed development and maturation. The results contradict the hypothesis that seed quality is maximal at the end of the seed-filling phase and that viability and vigour begin to decline immediately thereafter.     

施用纳米碳对烤烟氮素吸收和利用的影响

为明确纳米碳在提高烤烟氮素吸收利用方面的效果,在盆栽条件下,研究了纳米碳不同用量对烤烟根系生长发育、干物质积累和氮素吸收利用的影响。结果表明,在常规肥料中添加纳米碳能够促进烤烟根系生长发育,明显提高烟株根系活力和单株根系生物量,增加植株干物质积累量。施用纳米碳增加了烤烟植株成熟期各器官氮素含量和积累量,而未明显影响氮素在植株不同器官的分配。施用纳米碳不仅增加了植株对肥料氮的吸收量,还增加了对土壤氮的吸收量,这与其促进烤烟根系生长发育、提高根系吸收能力有密切关系。纳米碳无论做基肥还是做追肥,均显著提高了氮肥利用率,提高幅度分别达到14.44%和9.62%,有效降低了氮素土壤残留和损失。     

MG2+, K+-DEPENDENT PHOSPHATASE ACTIVITY DURING SEED DEVELOPMENT IN NICOTIANA TABACUM

Cation-dependent phosphatase activity was followed in developing Nicotiana tabacum (tobacco) seeds from pre-pollination to seed maturity. Enzyme activity increased rapidly up to 6 days post-pollination, then dropped off until 13 days after pollination. At 15 days post-pollination, enzyme activity had increased again, and at 17 days post-pollination, it reached its highest level. At 19 days post-pollination (seed maturity) enzyme activity was still 28% higher than that of the pre-pollination stage. Since the two peaks in enzyme activity correlate very highly with endosperm growth (first peak) and embryo growth (second peak), it is suggested that the Mg²⁺, K⁺-dependent phosphatase activity detected in this study is associated with the active transport of nutrients to the endosperm and embryo during seed development. The relatively high amount of enzyme activity remaining in the mature seed may represent a store of transport phosphatase that will be available for immediate use at the time of germination and early seedling growth.     

Seed Longevity of Rice Cultivars and Strategies for their Conservation in Genebanks

Changes in seed quality during ripening were studied in sixteencultivars of rice, representing the three ecogeographic racesofOryza sativa, and one cultivar ofO. glaberrima, grown duringone dry season (Nov. –May) 1992 –1993 at Los Baños, Philippines. Mass maturity (defined as the end ofseed filling period) among the cultivars was attained between18.5 and 21.6d after anthesis (DAA). The seed moisture contentat mass maturity varied between 24 and 40%. Germination abilityof seeds in the early stages of development varied significantly,but as mass maturity approached, germination increased to themaximum and no significant differences were found among cultivars.The seeds were stored hermetically at 35 °C with 15±0.2%moisture content and the resultant seed survival data were analysedby probit analysis. Potential longevity (quantified by the valueof seed lot constantK_iof the seed viability equation) was greatestbetween 33 and 37 DAA, i.e. about 2 weeks after mass maturity.The stage during development at which seeds achieve maximumpotential longevity is described by the term storage maturity.Lowlandjaponicacultivars, large seeded accessions (seed mass40mg) andO. glaberrimahad shorter storage longevity ( , standarddeviation of the frequency of seed deaths in time=1.47 weeks)while cultivars with purple pericarp survived longer than othercultivars ( =2.33 weeks). The initial germination of thejaponicacultivarsat storage maturity was high (99 –100%) and the estimatesof maximum potential longevity (K_i) which ranged between 3.3(Shuang cheng nuo) and 4.4 (Minehikare) were close to thoseof theindicacultivars. This research suggests that seed production environment betweenNov. and May at Los Ba ños is benign for the temperatejaponicacultivars.The implications of these results on management of rice geneticresources are discussed. Oryza sativaL.; rice; germplasm conservation; seed production environment; seed development; seed longevity     

Seed Production Environment, Time of Harvest, and the Potential Longevity of Seeds of Three Cultivars of Rice (Oryza sativa L.)

Changes in seed quality (assessed by potential longevity, i.e.the value of the seed lot constant K₁ of the seed viabilityequation) in three contrasting cultivars of rice (Oryza sativaL.) were monitored during seed development and maturation intwo temperature regimes, viz 28/20°C and 32/24°C (12/12h), provided by controlled environments. Mass maturity (definedas the end of the seed-filling phase) varied only between 18and 20 d after 50% anthesis. In five of the six treatment combinationsmaximum potential longevity was not achieved until 12-19 d aftermass maturity. In contrast, the maximum potential longevityof seeds of a japonica rice cultivar produced in the warmerregime was obtained in the first harvest after mass maturity.After mass maturity, the potential longevity of the japonicarice seed lots produced in the warmer environment was much lessthan that for the cooler environments. Maximum potential longevitywas also consistently greater in the cooler than the warmerregime for the two indica cultivars, although the differencein K₁ was small (0·3-0·5). The deleterious effectof increase in temperature on seed quality development was notdetected until after mass maturity. Maximum potential longevityin the cooler regime was greatest in the glutinous indica (K₁= 3·9) and least in the japonica cultivar (K₁ = 3·1).It is concluded that the japonica cultivar is not as well adaptedto warm seed production regimes as the indica cultivars. Consequently,subject to confirmation, this research suggest that the seedproduction of japonica cultivars for long-term genetic conservationshould be undertaken, whenever possible, in warm temperate environments.Copyright1993, 1999 Academic Press Oryza sativa L., rice, genebanks, seed development, seed storage, seed longevity, temperature     

Variant maturity in seed structures of <Emphasis Type="Italic">Pinus albicaulis</Emphasis> (Engelm.) and <Emphasis Type="Italic">Pinus sibirica</Emphasis> (Du Tour): key to a soil seed bank,unusual among conifers?

The seeds of Cembrae pines are dispersed by nutcrackers (Genus Nucifraga), which cache seeds in soil during autumn. The dispersal and establishment of seedlings via this mutualistic relationship is highly successful. On the other hand, irregular quality of seed crops and lack of detailed knowledge on germination process of Cembrae pine seeds hamper effective seedling production in the nursery. Therefore we studied basic structures and maturity of whitebark pine (Pinus albicaulis Engelm.) and Siberian stone pine (Pinus sibirica Du Tour) seeds, as well as structural changes during a multi-step treatment of whitebark pine seeds, using field emission scanning electron microscopy, transmission electron microscopy and light microscopy. The most striking differences compared to many other conifer seeds were the solid surface structures, early structural differentiation of the embryo, clustering of the thin-walled megagametophyte cells, and great accumulation of starch in both the untreated and treated seeds. Protein bodies of the embryo were in early developmental stages, whereas in the megagametophyte their stages varied. The number, form and size of lipid bodies also varied within different parts of the seed, and lipids dissolved easily. Our results indicated that despite maturity of the seed coat and advanced differentiation of the embryo, the embryo and the megagametophyte were still immature. These morphological features and a notable proportion of storage reserves remaining in unstable form may, however, be advantageous for maintaining viability and reaching maturity within a soil seed bank. Well-controlled pre-treatment simulating natural conditions should result in improved germination.     

Response of Soybean Cultivars to Nematicidal Treatments of Soil Infested with Meloidogyne incognita

A comparison of untreated and nematicide-treated soil for soybean production revealed that Meloidogyne incognita hastened crop maturity and reduced plant ht, seed wt, and yield. Reductions of yield varied from 32-90% depending on cultivar susceptibility. DBCP was more consistent in increasing crop performance than organo-phosphale or oxime carbamate nematicides. Greatest yield increases were produced by nematicidal treatment of soils planted to soybean cultivars with the lowest susceptibility.     

The effect of temperature and CO2 on seed quality development in wheat (Triticum aestivum L.)

Winter wheat (Triticum aestivum L.) cv. Hereward was grown inthe field in two double-walled polyethylene-covered tunnelswithin each of which a temperature gradient was superimposedon diurnal and seasonal fluctuations in temperature. The meantemperature between anthesis and harvest maturity varied from14.3 to 18.4C among plots within these tunnels. The CO₂ concentrationwas controlled at different values in each tunnel; seasonalmean concentrations were 380 and 684 µmol CO₂ mol^–1air. Crops were also grown outside the tunnels at ambient temperaturesand CO₂. Samples of seeds were harvested sequentially from eachplot between anthesis and harvest maturity. Seed germinationand seed survival during subsequent air-dry storage were determinedfor each sample. The onset of both ability to germinate anddesiccation tolerance (ability to germinate after rapid desiccationto 10–15% moisture content and subsequent rehydration)coincided in all environments. Full germination capacity (>97%, determined at 10C) was reached 4–18 d before theend of the seed-filling phase (mass maturity) in most cases.There was little or no decline in germination capacity duringsubsequent seed development and maturation. Differences in seedquality were evident, however, throughout seed development andmaturation when seed survival curves during subsequent storagewere compared. Potential longevity in air-dry storage (assessedby the value K₁ of the seed viability equation) improved consistentlyboth before and after mass maturity. There was a significantpositive relation between the rate of increase in potentiallongevity (dK₁Idt) and temperature (the minimum temperaturefor seed quality development was 4.8 C), but neither CO₂ concentrationnor production within the polyethylene tunnels affected thisrelation. Key words: Wheat, Triticum aestivum L., seed development, seed longevity, carbon dioxide, temperature     

Dynamic distribution and the role of abscisic acid during seed development of a lady’s slipper orchid,Cypripedium formosanum

Background and Aims Although abscisic acid (ABA) is commonly recognized as a primary cause of seed dormancy, there is a lack of information on the role of ABA during orchid seed development. In order to address this issue, the localization and quantification of ABA were determined in developing seeds of Cypripedium formosanum.Methods The endogenous ABA profile of seeds was measured by enzyme-linked immunosorbent assay (ELISA). Temporal and spatial distributions of ABA in developing seeds were visualized by immunohistochemical staining with monoclonal ABA antibodies. Fluoridone was applied to test the causal relationship between ABA content and seed germinability.Key Results ABA content was low at the proembryo stage, then increased rapidly from 120 to 150 days after pollination (DAP), accompanied by a progressive decrease in water content and seed germination. Immunofluorescence signals indicated an increase in fluorescence over time from the proembryo stage to seed maturation. From immunogold labelling, gold particles could be seen within the cytoplasm of embryo-proper cells during the early stages of seed development. As seeds approached maturity, increased localization of gold particles was observed in the periplasmic space, the plasmalemma between embryo-proper cells, the surface wall of the embryo proper, and the inner walls of inner seed-coat cells. At maturity, gold particles were found mainly in the apoplast, such as the surface wall of the embryo proper, and the shrivelled inner and outer seed coats. Injection of fluoridone into capsules resulted in enhanced germination of mature seeds.Conclusions The results indicate that ABA is the key inhibitor of germination in C. formosanum. The distinct accumulation pattern of ABA suggests that it is synthesized in the cytosol of embryo cells during the early stages of seed development, and then exported to the apoplastic region of the cells for subsequent regulatory processes as seeds approach maturity.     

Chilling tolerance in <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> induced by seed priming with putrescine

Chilling stress is one of primary constraints to tobacco production in many parts of the world. The present study was conducted to induce chilling tolerance in tobacco by seed priming with putrescine (Put) in relation to physiological changes, using seeds from two tobacco varieties, MSk326 (chilling sensitive variety) and Honghuadajinyuan (HHDJY, chilling tolerant variety). Seed germination, seedling antioxidant enzyme activities and malondialdehyde (MDA) concentration, as well as polyamine concentration were determined under low temperature. During chilling stress at 11°C, seed priming with 0.01 mM Put for 48 h (Put0.01mM48 h) and seed priming with 0.1 mM Put for 48 h (Put0.1mM48 h) significantly increased germination percentage, germination index, seedling length and dry weight of both varieties compared to the controls without Put treatment. When seedlings of 4-leaf stage suffered a short chilling stress (5°C), Put 0.1 mM 48 h improved the activities of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX), increased endogenous Put, Spd and Spm concentration and decreased the MDA concentration. The results showed that Put priming treatments were available to enhance the chilling tolerance of tobacco seedlings. The optimal treatment of Put was Put0.1 mM48 h.     

Seed development and maturation in Phaseolus vulgaris II. Post-harvest longevity in air-dry storage

Changes in seed quality were monitored during the developmentand maturation of seeds of bean (Phaseolus vulgaris L.) in differentenvironments by determining the subsequent survival of seedsin airdry storage. The seed survival curves (percentage normalgermination plotted against period of storage) conformed tonegative cumulative normal distributions, and the same seedstorage environment (40 C with 14% moisture content) providedthe same estimate of the standard deviation of the frequencydistribution of seed deaths in time for all the seed lots harvestedat different times during development in the different environments(21.3 d and 20.9 d in 1993 and 1994, respectively). The potentiallongevity of developing seeds (quantified by the value of theseed lot constant of the seed viablity equation) continued toincrease after mass maturity (end of the seed-filling phase).Maximum potential longevity was attained 23 d (30/24 C, 1993),13 d (30/24 C, 1994), or 34 d (27/21 C, 1994) after mass maturity,by which time seeds had dried naturally to 17% (30/24 C, 1993),18% (30/24 C, 1994), and 16% (27/21 C, 1994) moisture content.Seed quality was greater in the cooler temperature seed productionregime because of an increased duration of seed quality improvement.The results show that maximum seed quality is not attained untilsome time after the end of the seed-filling phase in bean, andconfirm that the slow desiccation that occurs after ovule abscissionduring natural maturation drying is beneficial to the developmentof seed quality in bean. Key words: Bean, Phaseolus vulgaris L., seed development, seed longevity, seed storage     

Yield,development, and quality response of dual‐toxin Bt cotton to Helicoverpa spp. infestations in Australia

To verify current thresholds for Bollgard II^® cotton in Australia, the impact of Helicoverpa spp. (Lepidoptera: Noctuidae) larvae on yield, development, and quality under various infestation intensities and durations, and stages of growth, was tested using small plot field experiments over two seasons. Infestation with up to 80 eggs m^?1 of Helicoverpa armigera (Hübner) and Helicoverpa punctigera Wallengren showed that species, infestation level, and stage of growth had no significant effect on yields of seed‐cotton or lint and on maturity and fibre quality. The duration of infestation of white flowers with H. punctigera neonates (maximum of every day for up to 4 weeks) had no impact on the yield of seed‐cotton or lint, maturity, and fibre quality, but when 100% of flowers were infested (compared with 0 or 50%), seed‐cotton and lint yields were significantly reduced and maturity was delayed. Infestation with up to 18 medium H. armigera larvae m^?1 at several plant stages did not significantly affect yields of seed‐cotton and lint, maturity, and fibre quality. A heliocide spray applied on a commercial farm at the current threshold resulted in a significantly higher lint yield, compared with a farm where no spray was applied. In conclusion, Bollgard II^® cotton is highly resistant to Helicoverpa spp. infestation.     

Seed production of a subalpine Festuca nigrescens–Agrostis capillaris semi-natural grassland in the eastern Italian Alps

Studying the seed production of semi-natural grasslands helps to understand their regeneration processes and to evaluate the possibility of collecting seed useful for ecological restoration. In a subalpine Festuca nigrescens–Agrostis capillaris meadow, the development of the main grasses' seed production and the standing seed yield at meadow maturity were studied. The inflorescence size was in the lower part of the variation range reported in previous studies. The seed viability was average to high and durable. The biological floret site utilization was mostly above 50%. With 458 fertile stems m^? 2, the seeds produced (all seeds produced up to the time of the meadow maturity) by the main grasses were 21,000 m^? 2, of which 77% were of A. capillaris and 81% were viable. On a 3-year average, the standing seed yield (seeds present on the plants, i.e. produced minus shed seeds) at meadow maturity was 6953 seeds m^? 2. Amount and composition of the produced seeds were affected mainly by the variability among years of the fertile stems density of the perennial grasses and the annual hemiparasite Rhinanthus freynii. The percentage of produced seeds which formed the standing seed yield was affected mainly by the collecting date.     

Acquisition of physical dormancy and ontogeny of the micropyle--water-gap complex in developing seeds of Geranium carolinianum (Geraniaceae)

Background and Aims

The ‘hinged valve gap’ has been previously identified as the initial site of water entry (i.e. water gap) in physically dormant (PY) seeds of Geranium carolinianum (Geraniaceae). However, neither the ontogeny of the hinged valve gap nor acquisition of PY by seeds of Geraniaceae has been studied previously. The aims of the present study were to investigate the physiological events related to acquisition of PY and the ontogeny of the hinged valve gap and seed coat of G. carolinianum.

Methods

Seeds of G. carolinianum were studied from the ovule stage until dispersal. The developmental stages of acquisition of germinability, physiological maturity and PY were determined by seed measurement, germination and imbibition experiments using intact seeds and isolated embryos of both fresh and slow-dried seeds. Ontogeny of the seed coat and water gap was studied using light microscopy.

Key Results

Developing seeds achieved germinability, physiological maturity and PY on days 9, 14 and 20 after pollination (DAP), respectively. The critical moisture content of seeds on acquisition of PY was 11 %. Slow-drying caused the stage of acquisition of PY to shift from 20 to 13 DAP. Greater extent of cell division and differentiation at the micropyle, water gap and chalaza than at the rest of the seed coat resulted in particular anatomical features. Palisade and subpalisade cells of varying forms developed in these sites. A clear demarcation between the water gap and micropyle is not evident due to their close proximity.

Conclusions

Acquisition of PY in seeds of G. carolinianum occurs after physiological maturity and is triggered by maturation drying. The micropyle and water gap cannot be considered as two separate entities, and thus it is more appropriate to consider them together as a ‘micropyle–water-gap complex’.     

True potato seed quality

Summary Environmental influence on physiological factors that determine seed quality is discussed in relation to true potato seed (TPS). This review has been motivated by the need for high quality seed for the proper evaluation of TPS technology. The objectives of tuber production may not be in harmony with those required for the production of high quality TPS. The production of high quality TPS may be influenced by the stronger sink strength ability of fast developing tubers for assimilation of available nutrients. Earliness of tuber formation, which is needed in TPS progenitors, may tend to result in incomplete TPS development. Limiting conditions during seed development decrease the potential of the seed for field establishment. Seed vigor is the most important attribute of seed quality since it is essential for seedling performance under the adverse conditions commonly encountered during field development. The attractiveness of TPS technology for tropical areas, where potato production is limited by low-quality seed tuber availability, is decreased by the lack of seedling vigor and uniformity of seedlings derived from TPS. Current research indicates that considerable genetic improvement of TPS vigor and uniformity is possible. It is suggested that significant immediate improvement of TPS quality would result from agronomic techniques that reflect understanding of the physiological factors that influence the production and maintenance of high quality seed. Research areas needed to develop methodologies for TPS production with optimum expression of genotype are identified.Abbreviations TPS True potato seed - CIP International Potato Center, Lima, Peru - SD Short day - LD Long day - INIA Instituto Nacional de Investigaciones Agropecuarias, Chile - GA Gibberellic acid     

Studies on seeds. II. Origin and degradation of lipid vesicles in pea and bean cotyledons

At least two kinds of lipid vesicles are present in pea and bean cotyledons which can be recognized at seed maturity on the basis of whether they do or do not interassociate into lipid vesicle sheets. Those that do interassociate into sheets are also characterized by (a) their association with plastids or plasma membranes during dormancy, and (b) the unique transformation into flattened saccules that they undergo during the first few days of seed germination. These interassociated (or composite) lipid vesicles have been found in only a few seeds and may be restricted to certain classes of plants and/or certain states of cellular development. Lipid vesicle-to-saccule transformation is predominantly confined to the germinating seed. However, some lipid vesicle-derived saccules are already present in some cells even before the seed reaches maturity. These partially transformed vesicles and saccules remain unchanged over dormancy, and then resume their transformation when the seed is germinated. This suggests that some stages of seed germination are already underway before the seed reaches maturity and are only resumed at seed germination. The lipid vesicles that do not interassociate into sheets (i.e., the simple lipid vesicles) are present in all tissues at all states of cellular development. These vesicles do not undergo any conspicuous structural changes during development.     

The Novel Approach to Enhance Seed Security: Dual Anti-Counterfeiting Methods Applied on Tobacco Pelleted Seeds

Seed security is of prime importance for agriculture. To protect true seeds from being faked, more secure dual anti-counterfeiting technologies for tobacco (Nicotiana tabacum L.) pelleted seed were developed in this paper. Fluorescein (FR), rhodamine B (RB), and magnetic powder (MP) were used as anti-counterfeiting labels. According to their different properties and the special seed pelleting process, four dual-labeling treatments were conducted for two tobacco varieties, MS Yunyan85 (MSYY85) and Honghua Dajinyuan (HHDJY). Then the seed germination and seedling growth status were investigated, and the fluorescence in cracked pellets and developing seedlings was observed under different excitation lights. The results showed that FR, RB, and MP had no negative effects on the germination, seedling growth, and MDA content of the pelleted seeds, and even some treatments significantly enhanced seedling dry weight, vigor index, and shoot height in MS YY85, and increased SOD activity and chlorophyll content in HHDJY as compared to the control. In addition, the cotyledon tip of seedlings treated with FR and MP together represented bright green fluorescence under illumination of blue light (478 nm). And the seedling cotyledon vein treated with RB and MP together showed red fluorescence under green light (546 nm). All seeds pelleted with magnetic powder of proper concentration could be attracted by a magnet. Thus, it indicated that those new dual-labeling methods that fluorescent compound and magnetic powder simultaneously applied in the same seed pellets definitely improved anti-counterfeiting technology and enhanced the seed security. This technology will ensure that high quality seed will be used in the crop production.     

Tissue-specific expression of a wheat high molecular weight glutenin gene in transgenic tobacco.

The expression of a wheat genomic clone containing the entire coding sequence of the high molecular weight glutenin subunit 12 gene flanked by 2.6 kilobases of 5' and 1.5 kilobases of 3' sequences has been studied after introduction into tobacco. Seeds of different tobacco plants containing the full-length wheat genomic clone accumulated different amounts of intact high molecular weight glutenin subunit mRNA and of a polypeptide displaying the solubility, molecular weight, and antigenic properties of the high molecular weight glutenin subunit 12. The wheat protein accumulated without obvious degradation products and constituted up to approximately 0.1% of the total tobacco endosperm protein. Restriction fragments corresponding to 2.6 kilobases, 1.4 kilobases, and 433 base pairs of high molecular weight glutenin 5' upstream sequence were fused to the coding sequence of the chloramphenicol acetyltransferase (CAT) gene in the vector polyCATter and transferred into tobacco. Chloramphenicol acetyltransferase enzyme activity was detected only in the seed endosperm tissue of the transformed plants. It was detected in tobacco seeds 8 days after anthesis and persisted until seed maturity. It is concluded that 433 base pairs of high molecular weight glutenin upstream sequence are sufficient to confer endosperm-specific expression of this monocot gene in the dicot tobacco.