Changes in Activities of Enzymes of Nitrogen Metabolism in Seedcoats and Cotyledons during Embryo Development in Pea Seeds

In the seedcoats of developing pea seeds, the maximal activities of asparaginase (EC 3.5.1.1) and aspartate: α-ketoglutarate aminotransferase (EC 2.6.1.1) are attained early in development, before the embryo has expanded to fill the embryo sac. These two enzyme activities could account for the early absence of asparagine and aspartate from the fluid secreted by the seedcoats into the embryo sac.     

Nutritive Role of the Seedcoats during Embryo Development in Pisum sativum L

Structural and metabolic features of the seedcoats of the developing pea seed indicate that events in the cells of the seedcoats are of major importance in controlling the development of the embryo. Information has been obtained on the distribution of N and P constituents of the seedcoats, embryo sac liquid, and the cotyledons of the embryo, in relation to the changes in the activities of several enzymes: aminopeptidases, β-glucosidase, and acid phosphatase. The liquid contents of the embryo sac are considered to arise as a secretion from the tegmen. High concentrations of amino acids (about 0.3 molar), NH₄⁺ (about 0.1 molar), and orthophosphate (Pi) (up to 4 millimolar) were measured in this fluid. Since Pi was the only form of P present, the data confirm the possible function of some of the seedcoat acid phosphatase activity in the provision of Pi to the embryo.     

Zeatin Metabolism in Fruits of Phaseolus: Comparison between Embryos, Seedcoat, and Pod Tissues

Zeatin metabolites were isolated from seedcoats and pod tissues of Phaseolus vulgaris and P. lunatus. The differences observed previously between P. vulgaris and P. lunatus embryos, i.e. the formation of O-ribosyl derivatives in the former and O-glucosyl derivatives in the latter, could also be detected in seedcoats, although the levels of these metabolites were much lower and there was a concomitant increase of breakdown products (adenine, adenosine and AMP). Inner pod wall tissues of both genotypes metabolized zeatin at a slow rate and the major metabolite was the mononucleotide of zeatin. The array of metabolites recovered was not influenced by the extraction method (cold ethanol or modified Bieleski solution).     

Studies of the Surfaces of Desert Plant Seeds: I. Effect of Day Length upon Maturation of the Seedcoat of Ononis sicula Guss

Pods of Ononis sicula may contain three types of seeds, allwith embryos ready to germinate but differing in the time whichthey take to imbibe water: seeds which have matured during longdays take longer than those matured in short days. The scanningelectron microscope has provided evidence that the delay inimbibition by the ‘long day’ seeds is related tothe thickness, i.e. the degree of development, of the cuticlesof their seedcoats.     

The effect soaking pea seeds with or without seedcoats has on seedling growth

Pea seeds (Pisum sativum L. `Alaska') with intact seedcoats (WC) and with seedcoats removed (WOC) were soaked in distilled water for 24 hours at 20°. The water, containing the pea diffusate, was decanted after the second, fourth, sixth, eighth, twelfth, and twenty-fourth hour and analyzed for total nitrogen, α-amino nitrogen, carbohydrate, and total solute dry weight. The seeds were germinated at 20° in a 16 hour photoperiod of 300 foot candles. Stem lengths and dry weights of roots, shoots and cotyledons were determined after 4, 11, and 18 days of growth. WOC seeds imbibed more water than WC seeds during the 24 hour imbibition period. Diffusates from WOC seeds always contained more solute than diffusates from WC seeds. Maltose, glucose, and fructose were not detected in the early diffusates from WOC seeds but were found in WC seed diffusates at all times. Seedlings from WC seeds had longer stems than those from WOC seeds. The dry weight of stems and roots of WC seedlings was greater than those from WOC seedlings. The dry weight of cotyledons from 18 day-old WC seedlings was less than from WOC seedlings. Water absorption by WC seeds was slower than by WOC seeds. Removal of the seedcoat allowed rapid imbibition resulting in seed injury presumably because of the loss of solutes which included monosaccharides, disaccharides, amino acids, and other nitrogen containing compounds. These results are consistent with the hypothesis that rapid imbibition disrupts membrane organization leading to reduction of seedling growth.     

Separation of Protochlorophylls Esterified with Different Alcohols from Inner Seed Coats of Three Cucurbitaceae

Reversed-phase high-performance liquid chromatography was usedto separate protochlorophyllide esters isolated from inner seedcoats of three Cucurbitaceae. At least 17 protochlorophyllsesterified with different alcohols were separated on an octadecylsilica (ODS) column from purified pigments of pumpkin (Cucurbitamoschata) and balsam pear (Momordica charantica) seeds witha single elution using 100% methanol within 22 min. The separationwas done according to the numbers of carbon atoms in the esterifyingalcohol of the protochlorophyll molecule with high resolutionand reproducibility. Gas chromatographic analysis of the pigment-esterifyingalcohols confirmed the presence of a number of protochlorophyllsesterified with different alcohols. This method permits rapid,efficient and quantitative detection and identification of chlorophyllsin complex mixtures. (Received May 17, 1982; Accepted September 6, 1982)     

Effect of the Osmotic Environment on K+ and Mg2+ Release from the Seed Coat and Cotyledons of Developing Seeds of Vicia faba and Pisum sativum. Evidence for a Stimulation of Efflux from the Vacuole at High Cell Turgor

After removal of the embryo from developing seeds of Vicia fabaL. and Pisum sativum L., the ‘empty’ ovules werefilled with a substitute medium (pH 5.5) and the effect of theosmolality of the medium on K⁺ and Mg²⁺ release from the seedcoat was examined. In long-term experiments (12 h or longer),with both attached and detached seed coats, the rate of K⁺ andMg²⁺ release from seed coats filled with a solution withoutosmoticum was enhanced, in comparison with release from seedcoats filled with a solution containing 400 mol m     

Changes in Photosynthate Unloading from Perfused Seed Coats of Phaseolus vulgaris L. Induced by Osmoticum and Ethylenediaminetetraacetate (EDTA)

Photosynthate unloading in Phaseolus vulgaris L. seed coatswas studied by treating perfused seed coats with differing concentrationsof an osmoticum and ethylenediaminetetraacetate (EDTA). Largechanges in osmoticum concentration typically produced rapidchanges in efflux of unlabelled sugar and steady-state-labelled¹⁴C-photosynthate. Osmoticum-induced changes in photosynthateefflux were caused by phloem import stimulation at low cellturgor and net efflux stimulation by high cell turgor. Eventhough rapid changes in sugar and tracer efflux were often inducedby osmoticum treatments, the specific activity of sugar releasedfrom seed coats was not greatly affected by these treatmentsand was similar to the specific activity of sugar remainingin the seed coat after perfusion. Thus, tracer was transportedfrom the phloem throughout the seed coat sugar pool before itwas released to the apoplast. This result is most consistentwith symplastic phloem unloading throughout perfused seed coats,because apoplastic transport between cells within the seed coatwas blocked by perfusion. Photosynthate efflux was stimulatedby simultaneous treatment of seed coats with EDTA and differentconcentrations of an osmoticum; loss of photosynthate from seedcoats did not appear to be tissue-specific. Key words: Phaseolus vulgaris, seed coat, photosynthate unloading, turgor, EDTA     

Phloem unloading in soybean seed coats: dynamics and stability of efflux into attached ;empty ovules'

The time-course of sucrose efflux from attached seedcoats (having their embryos surgically removed) into aqueous traps placed in the `empty ovules' had three phases. The first phase lasted 10 minutes and probably was a period of apoplastic flushing. The second lasted 2 to 3 hours and is thought to be a phase of equilibration of seed coat symplast with the frequently refreshed liquid. The third phase of relatively steady efflux was postulated to reflect the continued import of sucrose from the plant, and hence to reflect the rate of sieve tube unloading. The average steady state efflux was equal under most conditions to the estimated rate of sucrose import. Efflux and import were unaffected by 150 millimolar osmoticum (mannitol or polyethylene glycol [molecular weight about 400]), by 0.5 millimolar CaCl<sub>2</sub>, or by pretreatments up to 20 minutes with p-chloromercuribenzenesulfonic acid (PCMBS); they were enhanced by 40 micromolar abscisic acid, 40 micromolar indoleacetic acid, 20 micromolar fusicoccin, and 1 millimolar dithiothreitol (DTT) and were inhibited by 100 micromolar KCN, by 0.03% H<sub>2</sub>O<sub>2</sub>, by 20 micromolar and 5 micromolar trifluoromethoxy (carbonyl cyamide) phenylhydrazone, by repeated 5 minutes per hour treatments with 5 millimolar PCMBS, and by 5 millimolar DTT. The `steady state' sucrose efflux was able to account for about half the rate of dry weight growth of the embryo, but stabilization of the system with <1 millimolar DTT taken together with other considerations is likely to give good correspondence between experimental unloading rates and in vivo growth rates.     

Variation among 41 Genotypes of Tomato (Lycopersicon esculentumMill.) for Crossability toL. peruvianum(L.) Mill.

Even with the aid of tissue culture, crosses betweenLycopersiconesculentum(E) andL. peruvianum(P) typically yield few progeny.To determine whether some E genotypes produce more progeny perfruit that others when crossed with P, 41 E genotypes were crossedwith pollen bulked from five P accessions. This first experiment(expt 1) was replicated over 2 years. In a second experiment(expt 2), differences among three genotypes each of E and P,and among individual plants within E genotypes were investigated.The E genotypes for expt 2 were chosen for relatively high andlow crossability based on results of expt 1. The P genotypesfor expt 2 were from different accessions than those used inexpt 1. For both experiments, the 15 largest ovules from eachripe fruit were cultured aseptically for 1 month. Out of 1228fruit, 753 hybrids were obtained. For expt 1, significant genotypeby year interactions were observed. Within each year, therewere significant differences among E genotypes for crossability.In expt 2, significant effects were found for E genotypes, butnot for interactions between E and P genotypes, P genotypes,nor plants within E genotypes. Moreover, general crossabilityfor E genotypes using bulked pollen (expt 1) was indicativeof general crossability with three P accessions not presentin the bulk (expt 2). Thus, selecting E genotypes of high crossabilityto P is the key to obtaining progeny for gene introgression.Rare production of ExP seed which was large and had brown seedcoats typical of E seed indicated strong selection pressureto maintain separate species, but gene exchange in nature maybe possible albeit at a low rate over long periods of time. Interspecific hybridization; Lycopersicon esculentum; Lycopersicon peruvianum; ovule culture; speciation     

Cellular pathway of photosynthate transport in coats of developing seed of Vicia faba L. and Phaseolus vulgaris L. II. Principal cellular site(s) of efflux

The cells responsible for the photosynthate efflux from coatsof developing seed of Vicia faba L. and Phaseolus vulgaris L.were elucidated using known properties of the efflux mechanism.Sensitivity of sucrose efflux to NEM and high potassium concentrationswas retained by seed-coat halves of Phaseolus following pectinaseremoval of the branch parenchyma cell layer. In contrast, removalof the thin-walled parenchyma transfer cell layer from Viciaseed-coat halves abolished this sensitivity. The membrane-impermeantthiol-binding fluorochrome, qBBr, selectively stained the surfaceof the thin-walled parenchyma transfer cells. This phenomenonwas inhibited by the slowly permeable sul-phydryl agent, PCMBS,indicating that the plasma membranes of these cells are enrichedin sulphydryl groups characteristic of membrance porter proteins.On the basis that carrier-mediated sucrose efflux from seedcoats appears to be proton coupled, the putative plasma membraneH+-ATPase was used as a marker for the cells responsible forcarrier-mediated photosynthate efflux. When seed-coat halveswere exposed briefly at pH 8.5 to the weak acid fluorochrome,SRG, the ground parenchyma and thin-walled parenchyma transfercell layers selectively accumulated the dye. The apparent lowpH environment in the walls of these cells that renders SRGmembrane permeant appeared to be maintained by a VAN-sensitiveproton pump. The findings with SRG were corroborated by thecyto-chemical localization of plasma membrane ATPase activityto the ground parenchyma and thin-walled parenchyma transfercells using precipitation of cerium phosphate. Together, ourobservations provide qualified support for the conclusion thatcarrier-mediated photosynthate efflux from coats of Phaseolusand Vicia seed is primarily restricted to the ground parenchymaand thin-walled parenchyma transfer cell layers, respectively. Key words: Ground parenchyma, Phaseolus vulgaris L., photosynthate efflux, seed coat, transfer cell, Vicia faba L.     

Tissue-Specific Expression of Cell Wall Proteins in Developing Soybean Tissues

Cell wall hydroxyproline-rich glycoproteins (HRGPs) and glycine-rich proteins (GRPs) were examined at the protein and at the mRNA levels in developing soybean tissues by tissue print immunoblots and RNA blots. In young soybean stems, HRGPs are expressed most heavily in cambium cells, in a few layers of cortex cells surrounding primary phloem, and in some parenchyma cells around the primary xylem, whereas GRPs are highly expressed in the primary xylem and also in the primary phloem. In older soybean stems, HRGP genes are expressed exclusively in cambium cells and GRP genes are most heavily expressed in newly differentiated secondary xylem cells. Similar expression patterns of HRGPs and of GRPs were found in soybean petioles, seedcoats, and young hypocotyls, and also in bean petioles and stems. HRGPs and GRPs become insolubilized in soybean stem cell walls. Three major HRGP mRNAs and two major GRP mRNAs accumulate in soybean stems. Soluble HRGPs are abundant in young hypocotyl apical regions and young root apical regions, whereas in hypocotyl and root mature regions, soluble HRGPs are found only in a few layers of cortex cells surrounding the vascular bundles. GRPs are specifically localized in primary xylem cell walls of young root. These results show that the gene expression of HRGPs and GRPs is developmentally regulated in a tissue-specific manner. In soybean tissues, HRGPs are most heavily expressed in meristematic cells and in some of those cells that may be under stress, whereas GRPs are expressed in all cells that are or are going to be lignified.     

Permeabilization of metabolites from biologically viable soybeans (Glycine max)

Chemical permeabilization has been widely studied for the release useful metabolites from many types of plant cells and tissues. In this study, the effect of 0-30% (v/v) of aqueous methanol solutions were used to permeabilize soybeans for the release of two isoflavonoids: daidzein and genistein. The release of these metabolites increases with increasing methanol concentrations. The amounts of daidzein and genistein released can increase up to 40- and 86-fold, respectively, when incubated in a 30% (v/v) methanol solution for 24 h compared with those incubated with water only. The effect of methanol on the release rates is primarily due to an increase in solubility of the stored daidzein and genistein (14- to 18-fold) inside the seeds, thus maximizing the concentration gradients for metabolite release. However, the viability of the seeds dropped with increase in methanol concentrations and the incubation time. The viability of soybeans (indicated by their ability to germinate) after permeabilization treatment with 0-20% (v/v) methanol solutions was maintained above 80% throughout the 24 h, whereas no seeds were found to be viable when 30% (v/v) methanol solution was used. The permeability coefficients (P) of daidzein and genistein were found to increase as the methanol concentration used was increased. These P values were estimated to range from 1.1 x 10(-)(9) to 1.9 x 10(-)(8) m/s and 1.0 x 10(-)(9) to 1.7 x 10(-)(8) m/s, respectively. The increase in P can be attributed primarily to an increase in the partition coefficient of the metabolites in the soybean seedcoats. An empirical correlation is proposed in which the log P values are described as a function of the metabolite molecular weights and the partition coefficients of the metabolites between octanol and water, K(oct/water), which was modified to include the effect of methanol present. Knowledge obtained from this study will help provide useful selection criteria for chemical permeabilization of plant tissues, such as seeds, with minimal loss in their viability.     

Cellular pathway of photosynthate transport in coats of developing seed of Vicia faba L. and Phaseolus vulgaris L. I. Extent of transport through the coat symplast

The extent of post-phloem solute transport through the coatsymplasts of developing seeds of Vicia faba L. and Phaseolusvulgaris L. was evaluated. For Vicia seed coats, the membrane-impermeantfluorochrome, CF, moved radially from the chalazal vein to reachthe chlorenchyma and thin-walled parenchyma transfer cell layers.Thereafter, the fluorochrome moved laterally in these two celllayers around the entire circumference of the seed coat. Transferof CF from the chalazal vein was inhibited by plasmolysis ofattached ‘empty’ seed coats. In contrast, the spreadof phloem imported CF was restricted to the ground parenchymaof Phaseolus seed coats. Fluorochrome loaded into the outermostground parenchyma cell layer was rendered immobile followingplasmolysis of excised seed-coat halves. Phloem-imported [¹⁴C]sucroseand the slowly membrane permeable sugar, L-[¹⁴C]glucose, werepartitioned identically between the vascular and non-vascularregions of intact Vicia seed coats. For ¹⁴C-photosynthates,these partitioning patterns in attached ‘empty’Vicia seed coats were unaffected by PCMBS, but inhibited byplasmolysis. Tissue autoradiographs of intact Phaseolus seedcoats demonstrated that a pulse of ¹⁴C-photosynthate moved fromthe veins to the grounds tissues. In excised Vicia seed coats,preloaded with ¹⁴C-photosynthates, the cellular distributionof residual ¹⁴C-label was unaffected by PCMBS. In contrast,PCMBS caused the ¹⁴C-photosynthate levels to be elevated inthe veins and ground parenchyma relative to the branch parenchymaof Phaseolusseed coat halves. Based on the above findings, itis concluded that the phloem of Vicia seed coats is interconnectedto two major symplastic domains; one comprises the chlorenchyma,the other the thin-walled parenchyma plus thin-walled parenchymatransfer cells. For Phaseolusseed coats, the phloem forms amajor symplastic domain with the ground parenchyma. Key words: Phaseolus vulgaris L, phloem unloading, photosynthate transport, seed coat, symplast, Vicia faba L     

Comparative Studies of Acid Glycosidases from Three Legumes

The major isoenzymes of -mannosidase (EC 3.2.1.24 [EC] ) and ß-galactosidase(ECf 3.2.1.23 [EC] ) have been separated from cotyledons of gardenpea, Pisum sativum L. (Vicieae), chick pea, Cicer arietinumL. (Cicereae), and cowpea, Vigna unguiculata (L.) Walp. (Phaseoleae).Some of their properties have been determined, including pHoptima, K_m values for p-nitrophenyl glycosidc substrates, andthe effects of several inhibitors. Swainsonine, an indolizidinealkaloid, was the most effective inhibitor of mannosidase 1,with I₃₀ values of 5.6 x 10^–8 M (cowpea), 1x 10^–7M (chick pea) and 2.9 x 19^–7 M (pea). The most effectiveinhibitor of ß-galactosidase 2 from all sources wasD-galactonic acid-1,4-lactonwe (-lactone), with K_i values rangingbetween 3.0 and 3.9x 10^–3 M. An inhibitor of the E. coliß-galactosidose, p-aminophenyl thio-ß-D-galactopyranoside,did not inhibit any of the legume ß-galctosidases;rather it enhanced the activites of the enzymes from chick peaand cowpea cotyledons. Etiolated hull and seed tissues frompea pods developing in darkness contained similar acid glycosidaseactivities to normal green tissues, thus the chloroplast isan unlikely location for ß-galactosidase 2. The majorß-galactosidasesdetected with an indigogenic substrate (5-bromo-4-chloro-3-indoxyl-ß-D-galactopyranoside)following gel electrophoresis of extracts from pea hull, seedcoats and cotyledons appeared to be different from ß-galactosidase2. Acid glycosidase, cotyledon, isoenzyme, -lactone, legume, swainsonine     

Development of a scoring method to identify important areas of plant diversity in Ireland

In the face of accelerating biodiversity loss it is more important than ever to identify important areas of biodiversity and target limited resources for conservation. We developed a method to identify areas of important plant diversity using known species’ distributions and evaluations of the species importance. We collated distribution records of vascular plants and developed a scoring method of spatial prioritisation to assign conservation value to the island of Ireland at the hectad scale (10 km × 10 km) and at the tetrad scale (2 km × 2 km) for two counties where sufficient data were available. Each plant species was assigned a species conservation value based on both its conservation status and distribution in Ireland. For each cell, the species conservation values within the cell were summed, thereby differentiating between areas of high and low conservation value across the landscape. Areas with high conservation value represent the most important areas for plant conservation.The protected area cover and the number of species present in these important areas were also examined by first defining threshold values using two different criteria. Species representation was high in the important areas; the identified important areas of plant diversity maintained high representation of species of conservation concern and achieved high species representation overall, requiring a low number of sites (<8%) to do so. The coincidence of protected areas and important areas for plant diversity was found to be low and while some important areas of plant diversity might benefit from the general protection afforded by these areas, our research highlights the need for conservation outside of protected areas.     

生物多样性重要区域识别——国外案例、国内研究进展

生物多样性丧失已经成为全球重大环境问题之一,重要区域或重要物种的识别是制定和实施保护计划的首要步骤,生物多样性保护的优先性研究成为保护生物学研究的焦点之一。优先保护的概念很早就被提出,保护国际(Conservation International,CI)一直倡导的热点地区途径受到国际社会的重视,生物多样性重要区域(KBAs)可以是综合的,也可以是单一类群的重要区域,如不同的国家已经开展了鸟类重要区域(important bird areas,IBAs)、植物重要区域(important plants areas,IPAs)、蝴蝶重要区域(prime butterfly areas,PBAs)和两栖爬行动物重要区域(important amphibians and reptiles areas,IARAs)等的识别研究工作。集中力量优先保护一些重要的地区是目前生物多样性保护较为现实和高效的途径。以佛得角群岛(the Cape Verde Islands)、意大利(Italy)、荷兰(the Netherlands)分别依据动物、植物单一类群或多个类群组合进行生物多样性重要区域识别为例,介绍了几个国家的生物多样性重要区域识别经验,概述国内在生物多样性重要区域识别领域的研究现状,详细介绍了海南岛生物多样性保护优先区识别案例,同时以国务院2010年批准实施的《中国生物多样性保护战略与行动计划(2011—2030年)》划定的32个陆地生物多样性保护优先区为例,提出中国未来应全面开展生物多样性本底调查,在充分获取生物多样性分布数据的基础上,依据植被类型和物种多样性以及受威胁因素等,在32个陆地生物多样性保护优先区内进一步客观准确地识别生物多样性重要区域(热点中的热点或重要区域中的重要区域),为中国未来的保护地规划、生物多样性监测、政策制定等提供科学支撑。     

面向生态系统结构的生物多样性保护成效空间对比评估

开展生物多样性保护成效对比评估对于合理执行生态补偿机制的转移支付政策,以及进一步促进全国生态保护管理水平的有效提升都具有重要意义。以2015年版《全国生态功能区划》划定的生物多样性保护重要生态功能区为研究对象,采用生物多样性保护成效结构指数,探讨重要生态功能区生物多样性保护成效的区域分异情况。研究结果表明：(1)华南、华东与华中地区重要生态功能区的生态结构参照基准较高,东北地区重要生态功能区居中,较差的重要生态功能区则位于西南和西北地区,说明了全国生态环境本底的区域分异性;(2)西北和西南地区重要生态功能区的保护成效结构指数较高,华中和华南地区居中,而东北和华东地区的保护成效结构指数较低,体现了2015年全国重要生态功能区生物多样性保护成效的空间差异情况;(3)从草原、森林与湿地生物多样性的保护成效结构指数来看,西北与西南地区、东北和西南地区及华东地区的重要生态功能区分别具有较大优势。上述研究结果反映了全国重要生态功能区生物多样性保护成效优劣的空间差异状况,可为政府部门健全生态补偿机制以及制定生态系统恢复方案提供决策参考。     

Fertility on the US-Mexico border

Using Bongaarts' model, the relative importance of the proximate determinants of fertility is explored in five populations on the US-Mexico border. For the groups closest to natural fertility (the two Mexican groups), lactation, use of contraception, and marriage all were moderately important in terms of their direct effect on fertility. For the group with lowest fertility (Anglo-American), contraceptive use was an important factor inhibiting fertility; marriage was important but not nearly as important as contraceptive use. For the two US Mexican-American groups, contraceptive use was an important intermediate variable, not as important as for Anglo-Americans, but more important than it was for the two populations in Mexico. The proportion married was a moderately important factor for the Mexican-American groups. For these five populations the principal differences in fertility rates result from substantial differences in the use of effective contraception. Bongaarts' model proved very useful as an analytical framework in this study.     

Definition and spatial location of mouse interleukin-2 residues that interact with its heterotrimeric receptor.

The high affinity receptor for interleukin-2 (IL-2) contains three subunits called IL-2R alpha, beta and gamma. A biological and receptor binding analysis based on 1393 different mutant mouse IL-2 (mIL-2) proteins was used to define the function of each of the 149 residues. By this genetic analysis, 44 residues were assigned important functions, 21 of which were structural. The remaining 23 residues consisted of 19 residues, from three separate regions, that were important for IL-2R alpha interaction; three residues, from two separate regions, that were important for IL-2R beta interaction; and a single residue important for IL-2R gamma interaction. We built a model mIL-2 structure based on the homologous human IL-2 (hIL-2) crystal structure. The roles of the 21 residues presumed to be important for structure were consistent with the model. Despite discontinuity in the primary sequence, the residues specific for each IL-2R subunit interaction were clustered and located to three disparate regions of the tertiary mIL-2 structure. The relative spatial locations of these three surfaces are different from the two receptor binding sites known for the structurally related human growth hormone and the significance of this observation is discussed.