Molecular and biochemical analysis of the enzymes of cysteine biosynthesis in the plant <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Summary. Among the amino acids produced by plants cysteine plays a special role as a mediator between assimilatory sulfate reduction and provision of reduced sulfur for cell metabolism. Part of this characteristic feature is the presence of cysteine synthesis in plastids, mitochondria and cytosol. Plants are the major source of reduced sulfur for human and animal nutrition. Cysteine biosynthesis deserves special attention, since reduced sulfur is channelled from cysteine into many sulfur-containing compounds in food and feed. Recent investigations are reviewed that focus on structure and regulation of cysteine synthesis in the model plant Arabidopsis thaliana. These data indicate that cysteine synthesis is not just an intermediate reaction step but that it is part of a regulatory network that mediates between inorganic sulfur supply and the demand for reduced sulfur during plant growth and in response to environmental changes. Received December 3, 2001 Accepted December 21, 2001     

Molecular and biochemical analysis of serine acetyltransferase and cysteine synthase towards sulfur metabolic engineering in plants

Summary. Serine acetyltransferase (SATase) and cysteine synthase (O-acetylserine (thiol)-lyase) (CSase) are committed in the final step of cysteine biosynthesis. Six cDNA clones encoding SATase have been isolated from several plants, e.g. watermelon, spinach, Chinese chive and Arabidopsis thaliana. Feedback-inhibition pattern and subcellular localization of plant SATases were evaluated. Two types of SATase that differ in their sensitivity to the feedback inhibition by l-cysteine were found in plants. In Arabidopsis, cytosolic SATase was inhibited by l-cysteine at a physiological concentration in an allosteric manner, but the plastidic and mitochondrial forms were not subjected to this feedback regulation. These results suggest that the regulation of cysteine biosynthesis through feedback inhibition may differ depending on the subcellular compartment. The allosteric domain responsible for l-cysteine inhibition was characterized, using several SATase mutants. The single change of amino acid residue, glycine-277 to cysteine, in the C-terminal region of watermelon SATase caused a significant decrease of the feedback-inhibition sensitivity of watermelon SATase. We made the transgenic Arabidopsis overexpressing point-mutated watermelon SATase gene whose product was not inhibited by l-cysteine. The contents of OAS, cysteine, and glutathione in transgenic Arabidopsis were significantly increased as compared to the wild-type Arabidopsis. Transgenic tobacco (Nicotiana tabacum) (F₁) plants with enhanced CSase activities both in the cytosol and in the chloroplasts were generated by cross-fertilization of two transgenic tobacco expressing either cytosolic CSase or chloroplastic CSase. Upon fumigation with 0.1 μL L⁻¹ sulfur dioxide, both the cysteine and glutathione contents in leaves of F₁ plants were increased significantly, but not in leaves of non-transformed control plants. These results indicated that both SATase and CSase play important roles in cysteine biosynthesis and its regulation in plants. Received November 27, 2001 Accepted December 21, 2001     

Effect of dietary sulfur amino acids on the taurine content of rat tissues

Summary.  The effect of dietary sulfur amino acids on the taurine content of rat blood and tissues was investigated. Three types of diet were prepared for this study: a low-taurine diet (LTD), normal taurine diet (NTD; LTD + 0.5% Met), and high-taurine diet (HTD; LTD + 0.5% Met + 3% taurine). These diets had no differing effect on the growth of the rats. The concentration of taurine in the blood from the HTD- and NTD-fed rats was respectively 1,200% and 200% more than that from LTD-. In such rat tissues as the liver, the taurine content was significantly affected by dietary sulfur amino acids, resulting in a higher content with HTD and lower content with LTD. However, little or no effect on taurine content was apparent in the heart or eye. The activity for taurine uptake by the small intestine was not affected by dietary sulfur amino acids. The expression level of taurine transporter mRNA was altered only in the kidney under these dietary conditions: a higher expression level with LTD and lower expression level with HTD. Received January 8, 2002 Accepted January 18, 2002 Published online August 20, 2002 Authors' address: Dr. Hideo Satsu, Laboratory of Food Chemistry, Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan, Fax: +81-3-5841-8026 E-mail: asatsu@mail.ecc.u-tokyo.ac.jp Abbreviations: HTD, high-taurine diet; NTD, normal taurine diet; LTD, low-taurine diet; TAUT, taurine transporter; CSA, cysteine sulfinate; CDO, cysteine dioxygenase; CSAD, cysteine sulfinate decarboxylase; PBS, phosphate-buffered saline; DIDS, 4,4′-diisothiocyanostilbene-2′,2′-disulfonic acid     

Synthesis of novel acetylene-containing amino acids

Summary.  Novel synthetic procedures for the modification of non-proteinogenic acetylene-containing amino acids have been developed. The functionalization either proceeds via zinc/copper-mediated introduction of alkyl substituents, or via tungsten-catalyzed ring-closing alkyne metathesis reactions. Received March 28, 2002 Accepted October 3, 2002 Published online December 18, 2002 Acknowledgements These investigations are supported (in part) by the Netherlands Research Council for Chemical Sciences (CW) with financial aid from the Netherlands Technology Foundation (STW). Authors' address: Floris P. J. T. Rutjes, Prof. Dr., Department of Organic Chemistry, University of Nijmegen, Toernooiveld 1, NL-6525 ED Nijmegen, The Netherlands, E-mail: rutjes@sci.kun.nl  2, selected data: ¹H NMR (300 MHz, CDCl₃) δ 5.32 (d, J = 7.7 Hz, 1H), 4.44–4.40 (m, 1H), 3.76 (s, 3H), 2.75–2.73 (d, J = 5.0 Hz, 2H), 1.44 (s, 9H); ¹³C NMR (75 MHz, CDCl₃) δ 171.0, 155.0, 80.3, 74.6, 52.6, 51.9, 41.7, 28.3, 24.0; mp = 55°C.  Typical procedure for 5: zinc dust (116 mg, 1.408 mmol) was weighed into a 20 mL flask, which was repeatedly evacuated (with heating using a heat gun) and flushed with argon. Dry DMF (0.5 mL, distilled from CaH₂) and 1,2-dibromoethane (9.2 μL, 0.106 mmol) were added and the flask was heated at 80°C for 40 min. The reaction mixture was allowed to cool to room temperature, trimethylsilyl chloride (4 μL, 0.035 mmol) was added and the resulting mixture was stirred vigorously for a further 30 min under argon. Iodocyclohexane (69 μl, 0.528 mmol) was added and stirred at room temperature for 3 h more after which stirring was ceased to settle the zinc. CuCN (41 mg, 0.458 mmol) and LiCl (40 mg, 0.915 mmol) were heated to 150°C for 2 h and cooled to room temperature. Addition of DMF (1 mL) formed a soluble CuCN·2LiCl complex within 5 min. After cooling the Cu-complex to −15°C, the organozinc reagent was added dropwise followed by the bromoacetylene 2 (116 mg, 0.352 mmol). The mixture was allowed to stir overnight at room temperature. Water was added and the suspension was extracted using heptane, washed with brine, dried (MgSO₄) and concentrated. Purification using flash column chromatography (10% EtOAc in heptane) yielded 5 (100 mg, 81%) as a colorless oil. 5: IR ν 3355, 2929, 2852, 2359, 2337, 1749, 1717, 1498, 1447, 1365, 1251, 1181, 1060; ¹H NMR (300 MHz, CDCl₃) δ 5.28 (d, J = 7.7 Hz, 1H), 4.43–4.38 (m, 1H), 3.73 (s, 3H), 2.69–2.63 (m, 2H), 2.13 (m, 1H), 1.73–1.22 (m, 10H), 1.43 (s, 9H); ¹³C NMR (75 MHz, CDCl₃) δ 171.4, 155.0, 88.1, 79.9, 73.8, 52.3, 32.7, 32.7, 28.8, 28.2, 25.8, 24.6, 23.1; HRMS (EI): calculated for C₁₇H₂₇NO₄ 309.1940, found 309.1937.  A solution of the tungsten catalyst (7 mg, 10 mol%) in C₆H₅Cl (2 mL) was treated with a solution of 14 (49.0 mg, 0.120 mmol) in C₆H₅Cl (5.0 mL) under an argon atmosphere and the resulting mixture was heated at 80°C for 3 h. Evaporation followed by flash column chromatography (80% EtOAc in heptane) afforded 15 (21.0 mg, 50%; 64% after correction for starting material) and 14 (16 mg, 33%) as colorless oils. 15: [α]_D =–14.6 (c = 1, CH₂Cl₂); IR ν 3313, 2931, 2865, 2249, 1744, 1667, 1520, 1366, 1170; ¹H NMR (400 MHz, CDCl₃) δ 7.14 (d, J = 8.7 Hz, 1H), 6.08 (d, J = 8.3 Hz, 1H), 4.78 (q, J = 6.8 Hz, 1H), 4.27 (q, J = 7.9 Hz, 1H), 3.73 (s, 3H), 2.17–2.15 (m, 4H), 2.07–1.96 (m, 2H), 1.79–1.52 (m, 4H), 1.45 (s, 9H), 0.89–0.83 (m, 2H); ¹³C NMR (100 MHz, CDCl₃) δ 173.2, 171.8, 155.8, 80.4, 80.2, 79.3, 53.8, 52.5, 51.2, 32.8 (2×), 28.1, 24.6, 24.2, 18.3 (2×); HRMS (EI): calculated for C₁₈H₂₈N₂O₅     

Comparison of the in vitro and in vivo neurotoxicity of three new sources of kainic acid

Summary.  Historically, all commercially available kainic acid has been derived from a single biological source using a consistent method of extraction and purification. That source became unavailable in 1995. Recently, three new commercial suppliers of kainic acid have made the product available, but the source of the material and the purification processes used differ. Our objective was to systematically compare the response produced by each of these new sources of kainic acid using three established neurobiological techniques: neuronal cell culture, hippocampal slice electrophysiology, and whole animal behavioural toxicity. Results in all three systems indicated no overall differences between the three formulations, although studies in both cerebellar neuron cultures and whole animal toxicity testing in mice, revealed some significant differences that may imply subtle differences in receptor selectivity and/or potency. We conclude that all three sources of kainic acid are viable alternatives to traditional kainate but they may not be identical. Until further information becomes available researchers may want to avoid using the three formulations interchangeably, and take note of the source of kainic acid when evaluating literature describing results from other laboratories. Received June 29, 2001 Accepted August 6, 2001 Published online June 26, 2002     

Chromatographic determination of L- and D-amino acids in plants

Quantities of free L- and D-amino acids (L- and D-AAs) in plants (leaves of coniferous and decidious trees, fleshy fruits, leaf blades of fodder grasses, and seeds and seedlings of edible legumes) were determined. Amino acid (AA) enantiomers were converted into diastereomers using pre-column derivatization with o-phthaldialdehyde together with N-isobutyryl-L(or D)-cysteine followed by separation of the resulting fluorescent isoindol derivatives on an octadecylsilyl stationary phase using high-performance liquid chromatography. Relative amounts of D-AAs were also determined by enantioselective gas chromatography-mass spectrometry on Chirasil-L-Val. Free D-AAs acids in the range of about 0.2% up to 8% relative to the corresponding L-AAs acids were found in plants. D-Asp, D-Asn, D-Glu, D-Gln, D-Ser and D-Ala could be detected in most of the plants, and D-Pro, D-Val, D-Leu and D-Lys in certain plants. As D-AAs were detected in gymnosperms as well as mono- and dicotyledonous angiosperms of major plant families it is concluded that free D-AAs in the low percentage range are principle constituents of plants.     

A study on the estimation of sulfur-containing amino acid metabolism by the determination of urinary sulfate and taurine

Summary.  Sulfate and taurine are major end products of sulfur-containing amino acid metabolism in mammals including humans, and they are excreted in urine. Average excretions (μmol/mg of creatinine) in the morning urine of 58 female college students were: total (free plus ester) sulfate (a), 12.53 ± 3.85; free sulfate, 11.57 ± 3.69; taurine, 0.78 ± 0.53. Ratio of total sulfate and taurine was 10 : 0.6. Regression lines obtained by plotting total sulfate, free sulfate, or total sulfate plus taurine against urea have shown that the former excretions are significantly correlated with urea excretion. Excretion of total sulfate at zero point of urea excretion (b) was 5.30, which corresponded to 42.3% of average excretion (12.53) and was assumed to be derived from dietary sulfate. The difference 7.23 (a − b) seemed to be derived from sulfur-containing amino acids. It was pointed out that the difference of average sulfate excretion and sulfate excretion at zero urea excretion, namely a − b, was appropriate for the metabolic index of sulfur-containing amino acids of the group examined. As free sulfate constituted 92.3% of total sulfate, excretion of ester sulfate was at a constant level, and that of taurine was not significantly correlated with urea excretion, the value of free sulfate corresponding to the value a − b of total sulfate mentioned above seemed to be a reliable and convenient index in the assessment of sulfur-containing amino acid metabolism. Received December 3, 2001 Accepted January 2, 2002 Published online August 30, 2002 Authors' address: Dr. Toshihiko Ubuka, Department of Clinical Nutrition, Kawasaki University of Medical Welfare, Kurashiki Okayama, 701-0193 Japan, E-mail: ubukatos@mw.kawasaki-m.ac.jp     

A bacterial artificial chromosome library for soybean and identification of clones near a major cyst nematode resistance gene

 We constructed a bacterial artificial chromosome (BAC) library for soybean (Glycine max) consisting of approximately 30 000 clones with an average insert size of 120 kilobase pairs. The library was successfully screened with restriction fragment length polymorphism (RFLP) and microsatellite markers tightly linked to a major resistance gene for the cyst nematode, Heterodera glycines. Since many soybean RFLPs hybridize to duplicate loci, BACs homologous to duplicate RFLP loci were distinguished by digestion with the restriction enzyme originally used to map the RFLP, followed by a comparison of the hybridizing fragments. Linkage mapping of BAC clones identified with markers linked to the cyst nematode resistance gene demonstrated that these clones were located at the expected chromosomal positions and that there were no indications of chimeras within the genomic inserts. Received: 3 July 1997/Accepted: 26 August 1997     

Pyramiding of bacterial blight resistance genes in rice: marker-assisted selection using RFLP and PCR

 DNA marker-assisted selection was used to pyramid four bacterial blight resistance genes, Xa-4, xa-5, xa-13 and Xa-21. Breeding lines with two, three and four resistance genes were developed and tested for resistance to the bacterial blight pathogen (Xanthomonas oryzae pv. oryzae). The pyramid lines showed a wider spectrum and a higher level of resistance than lines with only a single gene. To speed up the gene pyramiding process and to facilitate future marker-aided selection, we developed PCR markers for the two recessive genes, xa-5 and xa-13, and used these to survey a range of rice germplasm. The results of the germplasm survey will be useful for the selection of parents in breeding programs aimed at transferring these bacterial blight resistance genes from one varietal background to another. Received: 6 December 1996/Accepted: 20 December 1996     

Genetic mapping in maize with hybrid progeny across testers and generations: plant height and flowering

DNA markers were used to identify quantitative trait loci (QTLs) for plant height, ear height, and three flowering traits in hybrid progeny of two generations (F_2:3, F_6:8) of lines from a Mo17×H99 maize population. For both generations, testcross (TC) progeny were developed by crossing the lines to three inbred testers (B91, A632, B73). The hybrid progeny from the two generations were evaluated at the same locations but in different years as per an early generation testing program. QTLs were identified within each TC population and for mean testcross (MTC) performance. Overall, more QTLs were detected in the F_6:8 than the F_2:3 generation. Totalled over all five traits, 41 (B91) to 69% (B73) of the QTLs for tester effects and 67% of the QTLs for MTC detected in the F_2:3 generation were verified in the F_6:8 generation. Although differences in relative rank of the QTL effects across generations were observed, especially for the flowering traits, parental contributions were nearly always consistent. Several (8–11) QTLs were identified with effects for all three tester populations and for all traits except the anthesis-silk interval, which had only two such regions. Over all five traits, previous evaluations in this population identified 26 QTLs with consistent effects for two (F_2:3, F_6:8) inbred-progeny evaluations, and 20 (77%) were also associated with MTC in at least one of the generations evaluated herein. In all instances of common inbred and TC QTLs, parental contributions were the same. Received: 26 November 1999 / Accepted: 18 April 2000     

Modified ELISA for the detection of neomycin phosphotransferase II in transformed plant species

 Identifying transformed plant lines carrying the antibiotic resistance marker gene, neomycin phosphotransferase II, requires a more definitive test than the ability of the plant to grow on kanamycin. Although a number of alternative assays have been described, most are cumbersome, time consuming and/or require the use of radioisotopes. This report describes an ELISA for the detection of the neomycin phosphotransferase II enzyme in transformed plant tissue. The ELISA utilises commercially available antibodies and provides a number of advantages, including an extremely low background, a reduction in the amount of tissue required for testing, and semi-quantitative data on neomycin phosphotransferase II gene expression. This method has been applied successfully to a number of independently transformed lines in nine plant species. Received: 4 January 1999 / Accepted: 17 April 1999     

Construction and characterization of a bacterial artificial chromosome library of peach

A peach [Prunus persica (L.) Batch] bacterial artificial chromosome (BAC) library of var. Jingyu was constructed. Jingyu is a traditional variety, that displays many of the important agronomic characters of stone fruits. Since peach leaves are rich in polysaccharides, high-molecular-weight (HMW) DNA was extracted from leaf nuclei using a protocol adapted to peach. The HMW DNA embedded in agarose plugs was partially digested by HindIII. After size-selection by pulsed field gel electrophoresis, the selected DNA fragments were ligated to pBeloBAC11 and transformed into E. coli DH10B cells by electroporation. In total 20,736 recombinant clones were obtained. The BAC library has an average insert size of 95 kb and represents approximately 6.7 peach haploid genome equivalents. The BAC clones were stable in E. coli cell after 100 generations. The lack of hybridization to chloroplast and mitochondrial genes demonstrated that the library is predominantly composed of nuclear DNA. The library was screened with two molecular markers, W4 and P20, that are linked to white flesh and nectarine genes of peach, respectively. Ten positive clones were detected. Their fingerprints will be used to determine clone relationships and assemble contigs. This library should be well-suited for the map-based cloning of peach genes and genome physical mapping. Received: 18 January 2000 / Accepted: 29 May 2000     

Cysteine metabolism in periportal and perivenous hepatocytes: Perivenous cells have greater capacity for glutathione production and taurine synthesis but not for cysteine catabolism

Summary.  Hepatocyte preparations highly enriched in cells from either the periportal or the perivenous zone of the liver acinus were prepared using a digitonin/collagenase perfusion method. Five enzymes of cysteine metabolism were assayed in both periportal and perivenous preparations. The ratios of periportal to perivenous activity were 0.76, 0.60, 0.81, 1.62, and 1.01 for cysteine dioxygenase, cysteinesulfinate decarboxylase, γ-glutamylcysteine synthetase, cystathionase, and asparate (cysteinesulfinate) aminotransferase, respectively. Only cysteinesulfinate decarboxylase activity was significantly different between periportal and perivenous cells. In incubations with 2 mmol/L [³⁵S]cysteine, total cysteine catabolism ([³⁵S]taurine plus [³⁵S]sulfate) between periportal and perivenous cells was not different, which is consistent with the observation of similar cysteine dioxygenase activity across the hepatic acinus. Consistent with the lower cysteinesulfinate decarboxylase activity in periportal cells, 16% of the total catabolism of [³⁵S]cysteine in periportal cells resulted in taurine synthesis compared to 28% in perivenous cells. A lower rate of [³⁵S]glutathione synthesis was observed in periportal cells compared to perivenous cells, but γ-glutamylcysteine synthetase activity was not significantly different between perivenous and periportal cells. Cysteinesulfnate decarboxylase can be added to the list of enzymes whose activities are markedly enriched in perivenous cells. Received January 15, 2002 Accepted February 4, 2002 Published online September 4, 2002 Acknowledgements This work was supported by the National Research Initiative Competitive Grants Program/United States Department of Agriculture Competitive Research Grant 02-37200-7583. Authors' address: Dr. Martha H. Stipanuk, Division of Nutritional Sciences, 227 Savage Hall, Cornell University, Ithaca, NY 14853-6301, U.S.A., E-mail: mhs6@cornell.edu     

A history of plant biotechnology: from the Cell Theory of Schleiden and Schwann to biotech crops

Plant biotechnology is founded on the principles of cellular totipotency and genetic transformation, which can be traced back to the Cell Theory of Matthias Jakob Schleiden and Theodor Schwann, and the discovery of genetic transformation in bacteria by Frederick Griffith, respectively. On the 25th anniversary of the genetic transformation of plants, this review provides a historical account of the evolution of the theoretical concepts and experimental strategies that led to the production and commercialization of biotech (transformed or transgenic) plants expressing many useful genes, and emphasizes the beneficial effects of plant biotechnology on food security, human health, the environment, and conservation of biodiversity. In so doing, it celebrates and pays tribute to the contributions of scores of scientists who laid the foundation of modern plant biotechnology by their bold and unconventional thinking and experimentation. It highlights also the many important lessons to be learnt from the fascinating history of plant biotechnology, the significance of history in science teaching and research, and warns against the danger of the growing trends of ignoring history and historical illiteracy.     

Plasma levels of amino acids in elderly long term care residents with oropharyngeal dysphagia: Comparison of hand-oral with tube-enteral-fed patients

Summary. Background: Dysphagia and eating difficulties are highly prevalent in long term care patients. Evaluation of their nutritional status is complicated by comorbidity, frailty and individual patterns of feeding. In previous studies we found vitamin deficiencies (folic acid B6 and B12) in orally fed elderly in early stages of oropharyngeal dysphagia despite satisfactory nutritional parameters (BMI, albumin and hemoglobin). The aim of this study is to evaluate the plasma amino acids levels in these hand-oral fed elderly patients with dysphagia. Methods: Plasma amino acids were measured in 15 orally fed elderly patients in early functional outcome swallowing scale (FOSS), stage 2, and compared with those of 15 matched nasogastric-tube-fed counterparts. Results: The plasma levels of all measured amino acids, ratio of essential to nonessential, levels of conditionally essential and the immune-enhancing amino acids were similar in both groups and within the normal range of our laboratory. The traditional nutritional parameters were also similar in both groups and within the normal range. Conclusions: Plasma levels of amino acids in elderly patients in early stage of FOSS are satisfactory, supporting the view that their protein intake is adequate. Further studies should concentrate on patients in advanced stages of FOSS.     

Genetic validation and characterization of RAPD markers differentiating black and red spruces: molecular certification of spruce trees and hybrids

 Picea mariana (black spruce) and P. rubens (red spruce) are closely related species which are difficult to differentiate morphologically. RAPD markers differentiating black and red spruces have been previously identified. In the present study, genetic validity of these markers was determined using samples representing range–wide provenances. Their applicability for certifying genetic identity of individual black, red trees and their hybrids from several sympatric and allopatric locations was demonstrated. These diagnostic fragments of both red and black spruce were present at a frequency of over 0.95 in allopatric provenances, but at a lower frequency in some sympatric provenances (0.43–1.00). Natural populations of red spruce exhibiting typical red spruce phenotype contained black spruce diagnostic RAPD fragments and black spruces growing in bogs with typical bog black spruce morphology, contained red spruce-specific RAPD markers. Some major RAPD markers were cloned and sequenced. The results reveal an extremely high degree of identity between the random primer and the primer binding sites on the genome. Amplification of black and red spruce genomic DNA with designed primers flanking the species-diagnostic RAPD markers indicates that most of RAPD markers used to differentiate black spruce from red spruce are not species specific since these sequences were detected in several spruce species using a more sensitive detection method. Received June 17, 2002; accepted August 5, 2002 Published online: February 4, 2003     

Determining the growing season of land vegetation on the basis of plant phenology and satellite data in Northern China

The objectives of this study are to explore the relationships between plant phenology and satellite-sensor-derived measures of greenness, and to advance a new procedure for determining the growing season of land vegetation at the regional scale. Three phenological stations were selected as sample sites to represent different climatic zones and vegetation types in northern China. The mixed data set consists of occurrence dates of all observed phenophases for 50–70 kinds of trees and shrubs from 1983 to 1988. Using these data, we calculated the cumulative frequency of phenophases in every 5-day period (pentad) throughout each year, and also drew the cumulative frequency distribution curve for all station-years, in order to reveal the typical seasonal characteristics of these plant communities. The growing season was set as the time interval between 5% and 95% of the phenological cumulative frequency. Average lengths of the growing season varied between 188 days in the northern, to 259 days in the southern part of the research region. The beginning and end dates of the surface growing season were then applied each year as time thresholds, to determine the corresponding 10-day peak greenness values from normalized difference vegetation index curves for 8-km² pixels overlying the phenological stations. Our results show that, at the beginning of the growing season, the largest average greenness value occurs in the southern part, then in the northern, and finally the middle part of the research region. In contrast, at the end of the growing season, the largest average greenness value is measured in the northern part, next in the middle and lastly the southern part of the research region. In future studies, these derived NDVI thresholds can be applied to determine the growing season of similar plant communities at other sites, which lack surface phenological data. Received: 29 November 1999 / Revised: 14 March 2000 / Accepted: 15 March 2000     

The diffusion model for migration and selection in a plant population

 The diffusion approximation is derived for migration and selection at a multiallelic locus in a partially selfing plant population subdivided into a lattice of colonies. Generations are discrete and nonoverlapping; both pollen and seeds disperse. In the diffusion limit, the genotypic frequencies at each point are those determined at equilibrium by the local rate of selfing and allelic frequencies. If the drift and diffusion coefficients are taken as the appropriate linear combination of the corresponding coefficients for pollen and seeds, then the migration terms in the partial differential equation for the allelic frequencies have the standard form for a monoecious animal population. The selection term describes selection on the local genotypic frequencies. The boundary conditions and the unidimensional transition conditions for a geographical barrier and for coincident discontinuities in the carrying capacity and migration rate have the standard form. In the diallelic case, reparametrization renders the entire theory of clines and of the wave of advance of favorable alleles directly applicable to plant populations. Received 30 August 1995; received in revised form 23 February 1996