Ultrastructure and post-floral secretion of the pericarpial nectaries of Erythrina speciosa (Fabaceae)

Background and Aims

The occurrence of nectaries in fruits is restricted to a minority of plant families and consistent reports of their occurrence are not found associated with Fabaceae, mainly showing cellular details. The present study aims to describe the anatomical organization and ultrastructure of the pericarpial nectaries (PNs) in Erythrina speciosa, a bird-pollinated species, discussing functional aspects of these unusual structures.

Methods

Samples of floral buds, ovaries of flowers at anthesis and fruits at several developmental stages were fixed and processed by the usual methods for studies using light, and scanning and transmission electron microscopy. Nectar samples collected by filter paper wicks were subjected to chemical analysis using thin-layer chromatography.

Key Results

The PNs are distributed in isolation on the exocarp. Each PN is represented by a single hyaline trichome that consists of a basal cell at epidermal level, stalk cell(s) and a small secretory multicellular head. The apical stalk cell shows inner periclinal and anticlinal walls impregnated by lipids and lignin and has dense cytoplasm with a prevalence of mitochondria and endoplasmic reticulum. The secretory cells show voluminous nuclei and dense cytoplasm, which predominantly has dictyosomes, rough endoplasmic reticulum, plastids, mitochondria and free ribosomes. At the secretory stage the periplasmic space is prominent and contains secretion residues. Tests for sugar indicate the presence of non-reducing sugars in the secretory cells. Nectar samples from PNs contained sucrose, glucose and fructose.

Conclusions

The secretory stage of these PNs extends until fruit maturation and evidence suggests that the energetic source of nectar production is based on pericarp photosynthesis. Patrolling ants were seen foraging on fruits during all stages of fruit development, which suggests that the PNs mediate a symbiotic relationship between ants and plant, similar to the common role of many extrafloral nectaries.     

Notes on Thai and Indo-Chinese Phaseoleae (Leguminosae-Papilionoideae)

Four new species, Cruddasia craibii, Flemingia kradungensis, F. tiliacea and Pueraria maesenii are described and illustrated. The following new combinations are proposed: Cruddasia pinnata; C. laotica, Dolichos grahamianus; D. oxyphyllus; Dolichovigna pilosa; Erythrina stricta var. suberosa; Flemingia macrophylla var. sootepensis; Pueraria candollei var. mirifica; Shuteria suffulta var. sinensis and Vigna radiata var. grandiflora. Pueraria rigens is moved to the tribe Millettieae (= Mellettia rigens).     

Sphagnum growth under N saturation: interactive effects of water level and P or K fertilization

• Sphagnum biomass is a promising material that could be used as a substitute for peat in growing media and can be sustainably produced by converting existing drainage‐based peatland agriculture into wet, climate‐friendly agriculture (paludiculture). Our study focuses on yield maximization of Sphagnum as a crop.
• We tested the effects of three water level regimes and of phosphorus or potassium fertilization on the growth of four Sphagnum species (S. papillosum, S. palustre, S. fimbriatum, S. fallax). To simulate field conditions in Central and Western Europe we carried out a glasshouse experiment under nitrogen‐saturated conditions.
• A constant high water table (remaining at 2 cm below capitulum during growth) led to highest productivity for all tested species. Water table fluctuations between 2 and 9 cm below capitulum during growth and a water level 2 cm below capitulum at the start but falling relatively during plant growth led to significantly lower productivity. Fertilization had no effect on Sphagnum growth under conditions with high atmospheric deposition such as in NW Germany (38 kg N, 0.3 kg P, 7.6 kg K·ha^?1·year^?1).
• Large‐scale maximization of Sphagnum yields requires precise water management, with water tables just below the capitula and rising with Sphagnum growth. The nutrient load in large areas of Central and Western Europe from atmospheric deposition and irrigation water is high but, with an optimal water supply, does not hamper Sphagnum growth, at least not of regional provenances of Sphagnum.

     

Compensatory growth responses to clipping defoliation in Leymus chinensis (Poaceae) under nutrient addition and water deficiency conditions

Compensatory growth responses of Leymus chinensis, a dominant species in Inner Mongolia steppe, to clipping defoliation were evaluated in a pot-cultivated experiment under different nutrient (N and P) and water availability conditions. Leymus chinensis exhibited over-compensatory growth at the light and moderate clipping intensities (20% and 40% aerial mass removed) with a greater accumulated aboveground biomass, higher relative growth rate (RGR), more rhizomatic tillers and a stimulation of compensatory photosynthesis to the remnant leaves as compared with those of the unclipped plants. Intense clipping (80% aerial mass removed), which removed most of the aboveground tissues, greatly reduced the growth of aboveground biomass in comparison with that of the unclipped plants. Nitrogen addition only slightly improved the biomass production and RGR in light and moderately clipped plants, and it did not allow plants in the intense clipping condition to over-compensate. Phosphorus addition had no obvious influences on the growth and physiological responses to clipping defoliation. These results indicated that nutrient addition could not compensate for the negative effects of severe clipping on the defoliated grass. On the other hand, there were no distinct positive responses under water deficiency condition for L. chinensis at all clipping intensities with a significant reduction of aboveground and belowground biomass, lower RGR, fewer rhizomatic tillers, and a lower net photosynthetic rate than other wet treatments. Additionally, the chlorophyll contents of remnant leaves gradually increased with the increase of clipping intensities in each treatment. In conclusion, although L. chinensis could compensate for tissues removal by some morphological and physiological responses, intense clipping and drought can result in a significant decrease of biomass and growth rate, even under enriched nutrition conditions.     

植物根系和叶片生长对水分亏缺的原初反应

细胞扩张生长是植物受水分亏缺影响最敏感的生理过程之一。主要在对细胞水分导性、细胞壁特性和延伸组织中溶质传输结果分析的基础上 ,从细胞、组织和器官水平上对细胞扩展生长进行了探讨。根系和叶片细胞主要通过以下 2个过程来补偿水分胁迫的作用 :调节扩展生长需要的细胞临界膨压 ;溶质在延伸组织中的运移。此外 ,还探讨了植物根系和叶片生长对水分亏缺的生理适应机制     

Photosynthesis and growth of Erythrina variegata as affected by water stress and triacontanol

Erythrina variegata Lam. seedlings were grown under water stress (Ψ = -3.2 MPa) and subsequently sprayed with triacontanol (Tria). Water stress significantly reduced shoot growth rate, while roots continued to grow. Content of chlorophyll (Chl) a decreased more than that of Chl b. Water stress also reduced photosynthetic activity of chloroplasts as measured by Chl fluorescence induction. Stress effect was identified at the oxidation site of photosystem (PS) 2 prior to the hydroxylamine donating site and perhaps close to or after the diphenylcarbazide donating site. The loss of O₂ evolving thylakoid polypeptides (33, 23, 17 kDa) and the large (55 kDa) and small (15 kDa) subunits of ribulose-1,5-bisphosphate carboxylase (RuBPC) were found in water stressed seedlings. The reduction in RuBPC activity was accompanied by reduction of CO₂ fixation and stomatal conductance. All photosynthetic parameters were improved by Tria. This revised version was published online in August 2006 with corrections to the Cover Date.     

Assessment of wheat productivity responses and soil health dynamics under brackish ground water

The continuous use of brackish groundwater for irrigation is detrimental for soil and crop attributes. A three-year research study was designed for the wheat crop to assess the effects of brackish groundwater on crop yield and soil health under a surface irrigation system. Three sites were selected in different cropping zones of Pakistan. The treatments comprised of irrigation with moderately brackish water having 0.8, 1.3 & 2.7 dSm^?1 of salinity and canal water. The results indicated that EC, SAR, bicarbonates, Ca²⁺ and Mg²⁺ levels increased in the soil for consecutive years and this increase was more at site S3 followed by S2 and S1. As soil depth is concerned, the increase was more pronounced in upper layers of soil (0–15 cm) as compared to 15–30 cm depth. Growth and yield were also affected by the consecutive use of this water, the number of plants, plant height, the number of spikes per plant, and yield was reduced at all the three sites. However, the impact was less pronounced at the site S1 whereas S3 was the most affected one. Grain weight and dry matter weight were observed to be maximum at S1. Water productivity was also calculated for all the three sites. Maximum water productivity was observed at S1 followed by S2 & S3. It was concluded that the continuous use of brackish water would have an adverse effect on crop yield and subsequently, soil health is also affected by it significantly.     

Hydrotropism: root growth responses to water

The survival of terrestrial plants depends upon the capacity of roots to obtain water and nutrients from the soil. Directed growth of roots in relation to a gradient in moisture is called hydrotropism and begins in the root cap with the sensing of the moisture gradient. Even though the lack of sufficient water is the single-most important factor affecting world agriculture, there are surprisingly few studies on hydrotropism. Recent genetic analysis of hydrotropism in Arabidopsis has provided new insights about the mechanisms that the root cap uses to perceive and respond simultaneously to moisture and gravity signals. This knowledge might enable us to understand how the root cap processes environmental signals that are capable of regulating whole plant growth.     

Effect of soil strength on root growth under different water conditions

The objective of this study was to determine the effects of soil water and soil strength on root growth in situations where the individual effects of both of these factors were important. Three grain legumes were grown from pre-germinated seeds for five days on 50-mm compacted columns of two major soils of Sri Lanka. Four or five levels of bulk density (1.1 to 1.8 Mg.m^–3) and five or six levels of matric potential (–0.02 to–2.0 MPa) were used.Soil strength and matric potential effects on root growth were independently significant for most crop and soil combinations. Under high (wet) matric potential (>–0.77 MPa) soil conditions, the effect of soil water on root growth was evident only in its effect on soil strength. Bulk density had a significant effect on root growth independent of soil strength and matric potential in three cases.For all crops and soils, root penetration was 80% of the maximum or greater when the average soil strength (soil water not limiting) was 0.75 MPa or less, and when the average matric potential (soil strength not limiting) was –0.77 MPa or greater (wetter). Root penetration was 20% of the maximum or less when the soil strength was greater than 3.30 MPa (soil water not limiting), and when matric potential (soil strength not limiting) was less than –3.57 MPa. The use of pre-germinated seeds, which contained imbibed water, combined with a lack of water loss from the closed chambers containing the plants is the probable cause for the very low (–3.57 MPa) matric potential that allowed root growth at 20% of the maximum.     

不同土壤水分状况对杨树、沙棘水分关系及生长的影响

在盆栽条件下对杨树、沙棘进行了3种土壤水分处理,研究结果表明：沙棘的叶含水率在同一土壤水分下比杨树高、而水势低,说明沙棘叶的抗旱保水能力强于杨树;沙棘单叶的光合速率能够长时间维持在较高的水平,蒸腾速率却并非如此。杨树无论在生长初期,还是生长中期其单叶WUE均高于沙棘。以新生枝生物量／耗水总量计算的整体WUE在3种土壤水分条件下,杨树的WUE分别是沙棘的2．4倍、2．3倍、2倍,其差异达极显著水平,但在3种土壤水分条件下2个树种都以中度亏缺下的水分利用率最高。整个生长季节处在严重干旱下的杨树和沙棘的生长均受到显著影响,沙棘生长受干旱影响程度小于杨树,从整个生长趋势上看杨树适宜于良好水分下的生长,沙棘在适宜水分和中度亏缺下均可良好生长。     

Physiological and growth responses of Centaurea maculosa (Asteraceae) to root herbivory under varying levels of interspecific plant competition and soil nitrogen availability

Summary Centaurea maculosa seedlings were grown in pots to study the effects of root herbivory by Agapeta zoegana L. (Lep.: Cochylidae) and Cyphocleonus achates Fahr. (Col.: Curculionidae), grass competition and nitrogen shortage (each present or absent), using a full factorial design. The aims of the study were to analyse the impact of root herbivory on plant growth, resource allocation and physiological processes, and to test if these plant responses to herbivory were influenced by plant competition and nitrogen availability. The two root herbivores differed markedly in their impact on plant growth. While feeding by the moth A. zoegana in the root cortex had no effect on shoot and root mass, feeding by the weevil C. achates in the central vascular tissue greatly reduced shoot mass, but not root mass, leading to a reduced shoot/root ratio. The absence of significant effects of the two herbivores on root biomass, despite considerable consumption, indicates that compensatory root growth occurred. Competition with grass affected plant growth more than herbivory and nutrient status, resulting in reduced shoot and root growth, and number of leaves. Nitrogen shortage did not affect plant growth directly but greatly influenced the compensatory capacity of Centaurea maculosa to root herbivory. Under high nitrogen conditions, shoot biomass of plants infested by the weevil was reduced by 30% compared with uninfested plants. However, under poor nitrogen conditions a 63% reduction was observed compared with corresponding controls. Root herbivory was the most important stress factor affecting plant physiology. Besides a relative increase in biomass allocation to the roots, infested plants also showed a significant increase in nitrogen concentration in the roots and a concomitant reduction in leaf nitrogen concentration, reflecting a redirection of the nitrogen to the stronger sink. The level of fructans was greatly reduced in the roots after herbivore feeding. This is thought to be a consequence of their mobilisation to support compensatory root growth. A preliminary model linking the effects of these root herbivores to the physiological processes of C. maculosa is presented.     

不同土壤水分条件下容重对玉米生长的影响

用玉米作为实验材料。进行分根实验研究不同土壤水分条件下容重对玉米生长的影响,种子根平分在装有塿土的分隔的白铁皮桶中,土壤容重分4种处理：低容重(两边容重都为1．20g·cm^-3)、中容重(两边容重都为1．33g·cm^-3)、高容重(两边容重都为1．45g·cm^-3)和混合容重(一边为1．20g·cm^-3,另一边为1．45g·cm^-3),土壤水分控制在高基质势(-0．17MPa)和低基质势(-0．86MPa)两个水平,结果表明,当植株生长在紧实土壤或土壤基质势从-0．17MPa降到-0．86MPa时。根长、根干重和地上部干重都显著降低,并且地上部干重的降幅更大,紧实土壤使根长降低的同时还使根的直径增大,无论是容重增大还是土壤水分含量降低所引起的高土壤阻力都使叶片扩展速度降低和植株变小,生长在紧实土壤中的植株变小不仅是因为叶片扩展速度降低,同时是成熟叶片叶面积缩小的结果。然而,当植株生长在混合容重土壤中时,处在低容重土壤中的根系生长得到加强,补偿甚至超补偿高容重土壤中根系生长的不足,整个植株的生长状况与低容重土壤中生长的植株接近。     

Mating behavior of Diabrotica speciosa (Coleoptera: Chrysomelidae)

Diabrotica speciosa (Germar) is an economically important pest of Neotropical cultures and represents a quarantine risk for Neartic and Paleartic Regions. Despite its agricultural importance, few studies have been done on mating behavior and chemical communication, which has delayed the development of behavioral techniques for population management, such as the use of pheromone traps. In this study, we determined 1) the age at first mating; 2) diel rhythm of matings; 3) number of matings over 7 d; 4) the sequence of D. speciosa activities during premating, mating, and postmating; 5) the duration of each activity; and 6) response to male and female conspecific volatiles in Y-tube olfactometer. The first mating occurred between the third and seventh day after adult emergence and the majority of pairs mated on the fourth day after emergence. Pairs of D. speciosa showed a daily rhythm of mating with greater sexual activity between the end of the photophase and the first half of the scotophase. During the 7 d of observation, most pairs mated only once, although 30% mated two, three, or four times. In a Y-tube olfactometer, males were attracted by virgin females as well as by the volatile compounds emitted by females. Neither males nor their volatiles were attractive to either sex. Our observation provide information about mating behavior of D. speciosa, which will be useful in future research in chemical communication, such as identification of the pheromone and development of management techniques for this species using pheromone traps.     

Germination and growth responses of hybridizing Carpobrotus species (Aizoaceae) from coastal California to soil salinity

Germination, growth, and physiological responses of hybridizing Carpobrotus from coastal California to soil salinity were studied. Hybrids are presumably the result of hybridization and introgression between the exotic Carpobrotus edulis, a succulent perennial invading coastal habitats, and the native or long-naturalized C. chilensis. Germination responses were investigated at 0, 10, 20, and 50% seawater. Seedling growth and physiology were compared by irrigating seedlings with solutions of the same seawater concentrations and in low and high nutrients. Germination was inhibited in the presence of salt, but recovered after transferring the seeds to fresh water. Seeds exposed to salt had higher final germination rates than control. Growth of Carpobrotus was slightly enhanced by low seawater concentrations but reduced at high salinity at both nutrient regimes. Leaf cell sap osmolarity increased with increasing soil salinity, and taxa did not differ significantly in this physiological adjustment. Leaf carbon isotope ratios (∂¹³C) ranged from −28 to −22‰ and became less negative at higher salinities, indicating an improved water use efficiency in the seedlings at high salt concentrations. In addition, ∂¹³C values were generally less negative at high than at low nutrients. Differences among taxa were generally small. The results show that salinity affects both establishment and growth of hybridizing Carpobrotus. The overall weak species differences in salt tolerance indicate that the exotic C. edulis can occupy the same sites as C. chilensis in terms of salinity. The similarity of hybrids in their response to salinity suggests that they may contribute to the invasion by Carpobrotus.     

Water relations and growth responses of Uniola paniculata (sea oats) to soil moisture and water-table depth

Summary This study examined the water relations and growth responses of Uniola paniculata (sea oats) to (1) three watering regimes and (2) four controlled water-table depths. Uniola paniculata is frequently the dominant foredune grass along much of the southeastern Atlantic and Gulf coasts of the United States, but its distribution is limited in Louisiana. Throughout most of its range, U. paniculata tends to dominate and be well adapted to the most exposed areas of the dune where soil moisture is low. Dune elevations in Louisiana, however, rarely exceed 2 m, and as a result the depth to the water table is generally shallow. We hypothesized that if U. paniculata grows very near the water-table, as it may in Louisiana, it will display signs of water-logging stress. This study demonstrated that excessive soil moisture resulting from inundation or shallow water-table depth has a greater negative effect on plant growth than do low soil moisture conditions. Uniola paniculata's initial response to either drought or inundation was a reduction of leaf (stomatal) conductance and a concomitant decrease in leaf elongation. However, plants could recover from drought-induced leaf xylem pressures of less than-3.3 MPa, but prolonged inundation killed the plants. Waterlogging stress (manifested in significantly reduced leaf stomatal conductances and reduced biomass production) was observed in plants grown at 0.3 m above the water table. This stress was relieved, however, at an elevation of 0.9 m above the water table. As the elevation was increased from 0.9 to 2.7 m, there were no signs of drought stress nor a stimulation in growth due to lower soil moisture. We concluded that although U. paniculata's moisture-conserving traits adapt it well to the dune environment, this species can grow very well at an elevation of only 0.9 m above the water table. Field measurements of water-table depth in three Louisiana populations averaged about 1.3 m. Therefore, the observed limited distribution of U. paniculata along the Louisiana coast apparently cannot be explained by water-logging stress induced by the low dune elevations and the corresponding shallow water-table depth.     

Physiological and biochemical responses of photomorphogenic tomato mutants (cv. Micro-Tom) under water withholding

In addition to mediating photomorphogenesis, phytochromes are responsible for many abiotic stress responses, acting upon biochemical and molecular mechanisms of cell signaling. In this work, we measured the physiological and biochemical responses of phytochrome-mutant plants under water stress. In tomato (Solanum lycopersicum L.), the aurea mutant (au) is phytochrome-deficient and the high-pigment-1 mutant (hp1) has exaggerated light responses. We examined the effects of water withholding on water potential, leaf gas exchange, chlorophyll fluorescence, chloroplast pigment content and antioxidant enzyme activity in au and hp1 and their wild-type cultivar Micro-Tom (MT). Initial fluorescence and potential quantum efficiency of photosystem II (PSII) photochemistry were not affected by the treatment, but effective quantum yield of PSII, electron transport rate decreased and non-photochemical quenching increased significantly in MT. Under water withholding conditions, MT had higher malondialdehyde concentration than the mutants, but au had higher activities of catalase and ascorbate peroxidase compared to the other genotypes. The tolerance of mutants to the effects of water withholding may be explained by the higher activity of antioxidant enzymes in au and by a higher concentration of antioxidant compounds, such as carotenoids, in hp1.     

Reproductive Biology of Erythrina falcata (Fabaceae: Papilionoideae)1

Reproductive phenology, floral biology, degree of self‐incompatibility, and floral visitors of Erythrina falcata were studied in an Argentinean population. Flowering occurs during the dry season from late August to late October. Flower lifetime is 5–6 d. Phylogenetic studies indicate that E. falcata, together with E. fusca and E. crista‐galli, are included in a basal clade within Erythrina. Its phylogenetic position, floral morphology, and nectar characteristics suggest a hummingbird–passerine mixed pollination system. The flowers are nontubular, and the vexillum (the upper petal of the corolla) covers the other remaining floral parts until displaced by a visiting passerine (Icterus cayanensis) or a hummingbird (Amazilia chionogaster). Both birds act as pollen vectors. Bees were observed as occasional pollinators. Nectar production begins at anther dehiscence and coincides with maximum stigmatic receptivity. The base of the keel forms a secondary nectar reservoir. Controlled pollinations showed that this species is self‐incompatible, although a few fruits develop from selfing. Pollen:ovule ratio (43,200:7) is as expected for a xenogamous plant. Only 1 percent of the flowers set seeds under natural conditions. Possible explanations for the low reproductive success are discussed.     

Alkaloid diversity in eleven species of Ormosia and in Clathrotropis macrocarpa (Leguminosae-Papilionoideae)

Quinolizidine alkaloids were surveyed using gas chromatography (GC) and combined mass spectroscopy (MS) in bark or leaves of eleven species ofOrmosia Jackson as well as inClathrotropis macrocarpa Ducke from tropical forests in Latin America. A list of detected alkaloids, with their MS fragmentation patterns and GC retention indices, is given, as are the distribution of alkaloids among the samples and their detected concentrations. Alkaloid diversity inOrmosia was high, and the taxonomic distribution of specific alkaloids was not congruent with morphological taxonomic criteria: Of 143 alkaloids apparent in 14 samples, 65 were observed in only one sample, and no single alkaloid was common to all samples. Within a species, alkaloid concentrations were inconsistent and therefore cannot be used as a taxonomic character. This was shown for leaf alkaloid concentrations of two neighboring conspecific trees in the same habitat. Hemos analizado mediante cromatografia de gases (CG) y espectrometría de masas (EM) la composición en alcaloides quinolizidínicos de muestras de corteza y hojas de once especies deOrmosia Jackson y deClathrotropis macrocarpa Ducke, procedentes de las selvas de Latinoamérica. Presentamos una lista completa de los alcaloides detectados incluyendo sus fragmentos en EM y su índice de retención en CG. También presentamos la distribución de los alcaloides en las muestras y su concentración. La variedad de alcaloides era alta y su distribución por especies no se ajustó con criterios taxonómicos morfológicos: De los 143 alcaloides detectados en 14 muestras, 65 alcaloides estaban presentes en una sola muestra y ningún en todas las muestras. La concentración de los alcaloides en una especie no fue consistente y por lo tanto no puede utilizarse como carácter taxonómico. Esta conclusión proviene del análisis de la concentración de alcaloides en hojas de dos árboles de la misma especie en el mismo hábitat.     

Mineral composition and growth of Colorado spruce (Picea pungens) seedlings under calcareous soil conditions

Summary Field studies were conducted to assess the mineral nutrition and growth of Colorado spruce (Picea pungens Engln) seedlings (2–4 yr) from provenances selected for superior growth on calcareous prairie soils. Tissue nutrient concentrations and response to nitrogen were determined by use of foliar analysis and growth pattern studies. Soil conditions ranged from 7.6–7.8 for pH, 12–23% for total CaCO₃, and 5–6% for active CaCO₃. Foliage mineral composition showed relatively low phosphorus (0.09–0.15%) and high calcium (0.45–1.52%) assimilation. Seasonal growth and seedling response to added nitrogen was not adversely affected by the calcareous soil condition. Levels of nitrogen in the foliage required for optimum growth ranged from 1.5–2.0% and were similar to that of other conifer species.     

Deep and intermediate complex morphophysiological dormancy in seeds of Ferula gummosa (Apiaceae)

We tested the hypothesis that seeds of the monocarpic perennial Ferula gummosa from the Mediterranean area and central Asia have deep complex morphophysiological dormancy. We determined the water permeability of seeds, embryo morphology, temperature requirements for embryo growth and seed germination and responses of seeds to warm and cold stratification and to different concentrations of GA₃. The embryo has differentiated organs, but it is small (underdeveloped) and must grow inside the seed, reaching a critical embryo length, seed length ratio of 0.65–0.7, before the seed can germinate. Seeds required 9 weeks of cold stratification at <10°C for embryo growth, dormancy break and germination to occur. Thus, seeds have morphophysiological dormancy (MPD). Furthermore, GA₃ improved the germination percentage and rate at 5°C and promoted 20 and 5% germination of seeds incubated at 15 and 20°C, respectively. Thus, about 20% of the seeds had intermediate complex MPD. For the other seeds in the seed lot, cold stratification (5°C) was the only requirement for dormancy break and germination and GA₃ could not substitute for cold stratification. Thus, about 80% of the seeds had deep complex MPD.