Growth of In vitro banana (Musa SPP.) shoots under photomixotrophic and photoautotrophic conditions

Summary In vitro banana (Musa spp.) shoots were cultured under photomixotrophic (30 gl⁻¹ sucrose and 0.2 h⁻¹ number of air exchanges of culture vessels) and photoautotrophic (0 gl⁻¹ sucrose and 3.9 h⁻¹ number of air exchanges) conditions for 28 d in 370 cm³ Magenta boxes (GA7-type) containing 70 ml of half-strength Murashige and Skoog (MS) medium with 22.2 μM N⁶-benzyladenine (BA). The effects of varying CO₂ concentration (475 or 1340 μmol mol⁻¹) and light intensity (photosynthetic photon flux (PPF) of 100 or 200 μmol m⁻² s⁻¹) were investigated. Fresh and dry weights of banana shoots grown photomixotrophically were significantly greater on day 28 than those grown photoautotrophically. Photoautorophic shoots had a larger number of unfolded leaves and greater leaf area than photomixotrophic plants by days 14 and 28, regardless of CO₂ concentration. The shoot fresh and dry weights on day 14 in photoautotrophic conditions were significantly greater at PPF of 200 μmol m⁻² s⁻¹ than at 100 μmol m⁻² s⁻¹. The increase in net photosynthetic rate of photoautotrophic banana shoots was significant compared with photomixotrophic shoots. The multiplication ratio of in vitro banana shoots grown photoautotrophically in a 28-d culture period was the greatest at 100 μmol m⁻² s⁻¹ PPF and 475 μmol mol⁻¹ CO₂.     

Responses of the photosynthetic system and peroxidase activity to the rooting conditions of oak micropropagation

The effects of indole butyric acid (IBA) concentration and the duration of the plant growth regulator (PGR) treatment on the rooting ability, peroxidase level and photosynthetic activity of young Quercus robur L. plants were studied. Four-week-old oak shoots were transplanted to rooting media supplemented with 10 or 20 μM IBA. After 2, 3, 4, 5 or 7 days the shoots were transplanted again to a fresh, PGR-free medium. On the tenth day after transfer to rooting medium, the CO₂ fixation capacity, pigment content and peroxidase activity were measured. The photosynthetic parameters varied as a function of the time spent on medium containing PGR, showing maximum values in plants transplanted on the third to fourth day to PGR-free medium. The rooting percentage of these plants reached its maximum within two weeks. However, peroxidase activity was the highest in plants transferred later to PGR-free medium. The most pronounced stimulating effect on rooting was achieved with the higher initial IBA concentration followed by a transfer to PGR-free medium on the third to fourth day. These plants showed the highest vitality and the best rooting ability. This revised version was published online in June 2006 with corrections to the Cover Date.     

Stomatal characteristics and leaf anatomy of potato plantlets cultured in vitro under photoautotrophic and photomixotrophic conditions

Summary Potato plantlets (Solanum tuberosum L. cv. Benimaru) were cultured under photoautotrophic (without any sucrose in the nutrient medium and with enriched CO₂ and high photosynthetic photon flux) and photomixotrophic conditions (20 g 1⁻¹ sucrose in the medium). Leaf anatomy and stomatal characteristics of the leaves were studied in relation to stomatal size and density. Leaf diffusive resistance, transpiration rate, and wax content of the leaves were also investigated. In the photoautotrophic treatment, stomata behaved normally by closing in the dark and opening in the light. The stomatal density increased twofold compared to that of the photomixotrophic treatment. Relatively thick leaves and an organized palisade layer were observed and the epicuticulal wax content was remarkably higher in this treatment, i.e., seven times greater than that of photomixotrophic treatment. In general, higher diffusive resistance of the leaves was observed than under photomixotrophic conditions; also the resistance increased in darkness and decreased in the light. All these characteristics led the plantlets to have a normal and controlled transpiration rate, which was exceptionally high in the photomixotrophic treatment throughout the light and the dark period.     

以根为外植体建立大蒜的组织培养体系

以国内4个大蒜栽培品种为材料,建立了以根为外植体的再生体系。将蒜瓣去皮后灭菌消毒,萌发后选取苗龄为5~7 d的无菌苗的根接种到含不同激素配比的培养基中进行愈伤组织诱导,发现MS+2,4-D 1 mg/L+2ip 0.1 mg/L组合愈伤诱导效率最高,平均为56.06%;愈伤组织经过2~3次继代培养,选取胚性愈伤组织置于不同分化培养基上进行培养,2~3个月后可见小芽产生,分化培养基为MS+KT 1 mg/L时,植株再生效率最高,平均达到35.01 %。本研究建立了一种以根为外植体的高效的大蒜愈伤诱导和再生体系,为大蒜遗传转化体系的建立打下良好基础。     

Development of photoautotrophy and photoinhibition of Gardenia jasminoides plantlets during micropropagation

This paper reports on the fast fluorescence responses of Gardenia jasminoides Ellis plantlets, at two successive stages (shoot multiplication and root induction) of culture in vitro. We test whether plantlets in vitro suffer photoinhibition during culture and whether the degree of photoautotrophy of these mixotrophic plantlets has any effect on the extent of photoinhibitory impairment. In this regard the effects of different sucrose levels in the medium and PPFD during growth on the development of photoautotrophy and the extent of photoinhibition were evaluated. Plantlets were grown under low, intermediate, and high (50, 100, and 300 mol m^-2 s^-1) PPFD, and at 3 different sucrose concentrations (0.5, 1.5, and 3.0%, w/v) in the medium, during shoot multiplication. During root induction the same growth conditions were assayed except for the high PPFD. The development of photoautotrophy was assessed via the difference between the stable carbon isotope composition of sucrose used as heterotrophic carbon source and that of leaflets grown in vitro. Plantlets from root induction showed more developed photoautotrophy than those from shoot multiplication. For both stages the low-sucrose medium stimulated the photoautotrophy of plantlets in vitro. In addition, intermediate PPFD induced photoautotrophy during shoot multiplication. For plantlets of both culture stages at the lowest PPFD no photoinhibition occurred irrespective of the sucrose concentration in media. However, during the shoot multiplication stage chlorophyll fluorescence measurements showed a decrease in F _v /F _m and in t _1/2 as growing PPFD increased, indicating photoinhibitory damage. The decline of F _v /F _m was caused mostly by an increase in F _o , indicating the inactivation of PSII reaction centers. However plantlets growing under low sucrose showed reduced susceptibility to photoinhibition. During root induction, only plantlets cultured with high sucrose showed a decrease in F _v /F _m as PPFD increased, although t _1/2 remained unchanged. In this case, the decline of F _v /F _m was mostly due to a decrease in F _m , which indicates increased photoprotection rather than occurrence of photodamage. Therefore, growth in low-sucrose media had a protective effect on the resistance of PSII to light stress. In addition, plantlets were more resistant to photoinhibition during root induction than during shoot multiplication. Results suggest that increased photoautotrophy of plantlets reduces susceptibility to photoinhibition during gardenia culture in vitro.Abbreviations AP apparent photosynthesis - Chl total chlorophyll content - Chl a/b chlorophyll a-to-b ratio - Chl/Car total chlorophyll-to-carotenoids ratio - ¹³C ratio of ¹³C/¹²C relative to PeeDee belemnite standard - F _m maximum chlorophyll fluorescence - F _o fluorescence emission when all reaction centres are open and the photochemical quenching is minimal - F _v variable chlorophyll fluorescence (F _m -F _o ) - F _v /F _m the ratio of variable to maximum chlorophyll fluorescence, indicator photochemical efficiency of PSII - MS medium Murashige and Skoog (1962) medium - PPFD photosynthetic photon flux density - Rd dark respiration, t _1/2 the half-time of the increase from F _o to F _m - IAA indole butyric acid     

Net photosynthetic and transpiration rates in a chlorophyll-deficient isoline of soybean under well-watered and drought conditions

The gas exchange traits of wild type soybeans (cv. Clark) and a near-isogenic, chlorophyll-deficient line homozygous for the recessive allele y9 (y9y9) were compared under either well-watered or water-stress conditions. Mature leaves of y9 had a 65% lower chlorophyll content than wild type. However, the net photosynthetic rate (PN) of y9 leaves was only 20% lower than in the wild type, irrespective of water availability. Transpiration rates (E) were significantly higher in leaves of y9, compared to the wild type, either under well-watered or stress conditions. The higher E of y9 correlated with increased stomatal conductance, particularly in the abaxial epidermis, where more than 70% of the stomata were located. The combination of lower PN and increased E resulted in a significant decrease of water use efficiency in y9, at both water availability levels. The relative water content decreased in stressed leaves, much more in y9 than in wild type leaves, probably because of the higher E of the mutant line. This revised version was published online in June 2006 with corrections to the Cover Date.     

间作对氮调控玉米光合速率和光合氮利用效率的影响

研究间作后作物光合碳同化和光合氮利用效率(PNUE)对氮投入的响应, 对阐释间作产量优势的氮调控效应, 指导间作氮肥管理有重要意义。本研究设置玉米(Zea mays)单作、玉米间作两种种植模式的4个氮水平(N0, 0 kg·hm ^-2; N1, 125 kg·hm ^-2; N2, 250 kg·hm ^-2; N3, 375 kg·hm ^-2), 分析间作与施氮量对玉米叶片特征、光合参数、PNUE和产量的影响。结果表明: 与单作相比, 间作显著增加玉米叶片的叶干质量和比叶质量; 各施氮水平(除N3)下, 间作中靠近马铃薯(Solanum tuberosum)侧的玉米叶面积均显著高于单作玉米。单间作对比发现, 间作提高了玉米光饱和点和暗呼吸速率。单作、间作靠玉米侧(I-M)、间作靠马铃薯侧(I-P)的玉米PNUE均随施氮量增加而降低, 降幅以I-P最大; 施氮量低于250 kg·hm ^-2时, 相同施氮量下的玉米PNUE和净光合速率(P_n)均以I-P最高, I-M和单作次之。间作显著提高了玉米产量(土地当量比>1)。该研究中当施氮量≤250 kg·hm ^-2时, 间作I-P的玉米叶片P_n和PNUE显著提高可能是间作玉米产量提高的重要原因。     

Supplemental manganese improves the relative growth, net assimilation and photosynthetic rates of salt-stressed barley

Previous results in our laboratory indicated that a reduced Mn concentration in the leaves of barley was highly correlated with the reduced relative growth and net assimilation rates of salt-stressed plants. If Mn deficiency limits the growth of salt-stressed barley, then increasing leaf Mn concentrations should increase growth. In the present study, the effect of supplemental Mn on the growth of salt-stressed barley ( Hordeum vulgare L. cv. CM 72) was tested to determine if a salinity-induced Mn deficiency was limiting growth. Plants were salinized with 125 mol m⁻³ NaCl and 9.6 mol m⁻³ CaCl₂. Supplemental Mn was applied in 2 ways: 1) by increasing the Mn concentration in the solution culture and 2) by spraying Mn solutions directly onto the leaves. Growth was markedly inhibited at this salinity level. Dry matter production was increased 100% in salt-stressed plants treated with supplemental Mn to about 32% of the level of nonsalinized controls. The optimum solution culture concentration was 2.0 mmol m⁻³, and the optimum concentration applied to the leaves was 5.0 mol m⁻³. Supplemental Mn did not affect the growth of control plants. Further experiments showed that supplemental Mn increased Mn concentrations and uptake to the shoot. Supplemental Mn increased the relative growth rate of salt-stressed plants and this increase was attributed to an increase in the net assimilation rate; there were no significant effects on the leaf area ratio. Supplemental Mn also increased the net photosynthetic rate of salt-stressed plants. The data support the hypothesis that salinity induced a Mn deficiency in the shoot, which partially reduced photosynthetic rates and growth.     

Relative growth rates of leaves from soybean grown under drought-stressed and irrigated field conditions

Abstract. The relative growth rates and leaf area were graphed against leaf area, normalized with respect to final leaf area, to assess the applicability of the Lockhart cell wall expansion equation to soybean, Glycine max (L.) Merr., leaf development under field conditions. For leaves that had completed more than 20% of their growth, relative growth rates decreased linearly with an increase in the normalized leaf area, indicating that these leaves were undergoing strictly expansive growth. Drought stress significantly decreased the relative growth rate of these larger leaves. Small leaves which had completed less than 20% of their growth, were found to have highly variable relative growth rates. The large variability in relative growth rates indicated that the Lockhart cell wall expansion equation was inadequate to evaluate the growth of these young leaves. Drought stress had virtually no influence on the relative growth rates observed in the small leaves.     

Extracellular formation of 5-aminolevulinic acid by intact cells of the marine photosynthetic bacterium Rhodovulum sp. under various pH conditions

Extracellular formation of 5-aminolevulinic acid (ALA) by Rhodovulum sp. PS88 correlated with the consumption of the undissociated form of levulinic acid (LA) in an intact cell system. The concentration of the undissociated form of LA governed the extracellular formation of ALA at various culture pH values. This phenomenon might be caused by inhibition of ALA dehydratase by the undissociated form of LA after uptake into the cells as observed in Rhodobacter sphaeroides.     

Effects of 3,4-dihydroxybenzoic acid on tobacco (Nicotiana tabacum L.) cultured in vitro. Growth regulation in callus and organ cultures

ABSTRACT

The influence of 3,4-dihydroxybenzoic acid (protocatechuic acid), a naturally occurring benzoic acid derivative, on tobacco (Nicotiana tabacum L.) cell and tissue cultures was examined. The response to 0.1, 10 and 1000 µM 3,4-dihydroxybenzoic acid was tested with regards to cell proliferation in leaf explants, callus growth and shoot formation. Effects on shoot and root growth in micropropagated plants were also analysed. The highest concentration of 3,4-dihydroxybenzoic acid strongly inhibited the proliferation of leaf tissues, callus growth, shoot regeneration and root growth in micropropagated plants. On the contrary, the lowest concentration (0.1 µM) showed auxin-like activity by stimulating cell dedifferentiation, callus induction and rooting of leaf tissues. The presence of auxins and cytokinins in the media contrasted 3,4-dihydroxybenzoic acid inhibition of callus growth at all tested concentrations.     

Nutrient inflows into apple roots. II. Nitrate uptake rates measured on intact roots of mature trees under field conditions

Abstract The rates of uptake of nitrate-N per unit length; surface area and volume of root were measured in solution depletion experiments conducted in a root laboratory, using intact roots of two 4.5-year-old apple trees (Discovery/M.9 and Worcester Pearmain/M.9) at two different depths in the soil profile. In Discovery/M.9, NO₃^? uptake rate per unit root was constant over the 20-200 mmol m^?3 range of solution concentration. In Worcester/M.9, the uptake rate per unit root over the 200-150 mmol m^?3 range (corresponding to a ‘lag’ phase) was lower than that over 150-20 mmol m^?3. The uptake rates after the lag phase at depths of 46 and 104 cm were ca. 1.3 and 5.0 times greater than those in Discovery/M.9 at the 46 and 110 cm depths, respectively. The concentration below which net uptake was zero was ca. 1 mmolm^?3. In Discovery/M.9, the uptake rate per unit root at the 46cm depth was about 2.8 times that at 110 cm whereas in Worcester/M.9, the uptake rates at 46cm depth were about 1.8 and 1.4 times lower than those at 104cm over the solution concentration ranges 200-150 and 150-20 mmol m^?3, respectively. Only small differences were observed in uptake rates per unit root between 1400-1700 h, 2400-0400 h, and 0700-1100 h. For successive 5°C-increments in root temperature between 5 and 25° C, the nitrate uptake rate per unit root increased by 130, 10, 30 and 5%, respectively. A major change in the activation energy for nitrate uptake was observed at a transition temperature located between 5°and 10°C.     

Growth of various ecotypes ofLemna paucicostata in Japan under various temperature conditions,and their wintering forms

Lemna paucicostata in Japan is classified into 4 types, N-1, N-2, K and S types. S type strains which are distributed in southern Japan grew more rapidly than other strains at the temperatures of both 27 and 17C. With the extension of the culture period at 17 C to which the plants had been transferred from 25C, the growth rate increased greatly in S type strains, while it decreased in N-1 type strains which are distributed widely from Hokkaido to Kyushu, and particularly in those distributed in the northern area. S type strains survived winter mostly in the form of normal fronds under natural conditions at Kyoto (minimum temperature is lower than OC), while most of the fronds of N-1 type strains died after producing seed before winter. Thus S type strains are considered to have adapted to the climate of southern Japan so as to survive winter in the form of fronds, while N-1 type strains overwinter in the form of seed. N-2 type strains which are distributed in the northern part of the area adjacent to the Japan Sea produced turion-like fronds during winter under natural conditions, while none of the other strains did. The K type strain which was found only at the campus of Kyoto University overwintered only in the form of fronds which produce no seed.     

Differences in growth,pigment composition and photosynthetic rates of two phenotypes Microcystis aeruginosa strains under high and low iron conditions

This paper provided insight into the influence of iron on the growth of Microcystis aeruginosa strains related to different phenotypes of this species. In this research it was intended to compare the growth, pigment composition, photosynthetic efficiency and extracellular polysaccharides production of unicellular and colonial strains of M. aeruginosa. A significantly growth inhibition under iron-limited condition on unicellular M. aeruginosa was noted, whereas the colonial strain could maintain a steady growth along with the culture time. This observation was reconfirmed by the content of chlorophyll a. Compared with unicellular strain; the colonial strain exhibited a higher PSII maximum light energy transformation, photosynthetic oxygen evolution and extracellular polysaccharides (EPS) production in iron-limited condition. Further, in order to gain more information about the accessibility of iron in the two phenotypic Microcystis, we found the two strains could produce hydroxamate-type siderophores, the content of siderophores produced by the colonial strain was more than those in unicellular strain under the iron-limited condition. It was interpreted as an adaptation to the dilute environment. Our results demonstrated that the colonial phenotypes possessed stronger ability to endure iron-limited condition than unicellular strain by higher pigment contents, higher photosynthetic activities, higher EPS production and higher siderophores secretion. It might elucidate that the colonial M. aeruginosa bloom can sustain in eutrophic reservoirs and lakes.     

Growth regulation, plastid differentiation and the development of a photosynthetic system in cultured carrot root explants as influenced by exogenous sucrose and various phytohormones

Chromoplasts, which exist in the cells of freshly isolated carrot root explants, seemed to be transformed in thylakoid containing plastids, and chlorophyll formation was initiated if the explants were cultured in a liquid medium containing inositol and IAA as a hormonal supplement. This process was intensified when kinetin was also added, but no dependence on a sucrose supply could be found.A sucrose supply of 2% in conjunction with the combination of all three hormones, however, was needed to achieve maximal thylakoid formation including stacking in individual chloroplasts and for the very extensive chloroplast multiplication in explants growing with high cell division activity. It should be noted that the number of plastids per cell is strongly increased by the sucrose supplement which leads also to starch accumulation. However, no transformation into chloroplasts occurred without the hormonal stimulus.     

Radial and axial turgor pressure measurements in individual root cells of Mesembryanthemum crystallinum grown under various saline conditions

Abstract. Radial and axial turgor pressure profiles were measured with the pressure probe in untreated and salt-treated intact roots of Mesembryanthemum crystallinum. The microcapillary of the pressure probe was inserted step-wise into the root tissue 5, 25 and 50 mm away from the root cap. For evaluation of the data, only those recordings on a given root were used in which four discontinuous increases in turgor pressure occurred. These four turgor pressure increases could be related to the rhizodermal cells and to the cells in the three cortical layers. The measurements showed that a radial turgor pressure gradient of the same magnitude (directed from the third cortical layer to the external medium) existed along the root axis. The magnitude of this turgor pressure gradient decreased with increasing salinity (up to 400 mol m^-3 NaCl) in the growth medium. Addition of 10 mol m^-3 CaCl₂ to the 400 mol m^-3 NaCl medium partly reduced the salt-induced decrease in turgor pressure, but only in cells 25–50 mm away from the root tip. Combined with this effect, a small axial turgor pressure gradient was generated, therefore, in the cortex layers which was directed to the root tip. Measurements of the volumetric elastic modulus, ?, of the wall of the individual cells showed that the presence of salt considerably reduced the magnitude of this parameter and that addition of Ca²⁺ to the strongly saline medium partially diminished this decrease. This effect was strongest in cells 50 mm away from the root tip. The magnitude of ? of rhizodermal and cortical cells increased along the root axis both in untreated and in salt-treated roots. The ? value was significantly smaller for rhizodermal cells compared to the cortical cells, with the exception of cells 50 mm from the tip. In this tissue, rhizodermal and cortical cells exhibited nearly the same values. The decrease of the ?-values with salt and the increase along the root axis under the various growth conditions could be correlated with corresponding changes in cell volume. Diurnal changes in turgor pressure could not be detected in the individual root cells, with the notable exception of the rhizodermal and cortical cells located in the region 50 mm away from the root tip of the control plants. In these cells, an increase in turgor pressure was observed during the morning hours. Determination of the average osmotic pressure in tissue sections along the roots of control and salt-treated plants revealed that at 400 mol m^-3 NaCl the osmotic pressure gradient between the tissue and the medium is exo-directed, provided that the water is not (partly) immobilized.     

Characterization of photosynthetic events and associated changes in various clones of tea(camellia sinensis l) under low temperature conditions

Low temperature is one of the major environmental factors determining the growth rate ofCamellia sinensis (L.), and photosynthesis is one major metabolic process commonly used as a tool for identifying low temperature stress effects on plants. The Fv/Fm values at 6:30 a.m. (300-400 umol m^-2s^-1) did not vary much between the clones of tea plants. Further, when the light incidence increased at mid noon hours (1700-1800 μmol m^-2s^-1), the Fv/Fm values recorded a decline, irrespective of the clones. Of the 10 clones of tea plants under investigation, 3 clones, namely CRA-6017, TTL-6 and SMP-1, recorded a sharp decrease in the Fv/Fm ratio by 2 p.m. The malondialdehyde (MDA) levels in all the clones increased from 9 a.m. (1500-1700 μmol m^-2s^-1) to 2 p.m. and from 4:30 p.m. (900-1100 μmol m^-2s^-1) onwards it started to decrease and reach the levels equivalent to 6:30 a.m. by 7 p.m. (5–10 μmol m^-2s^-1 ). But the clones of TTL-1, TTL-4 and UPASI-9 showed low temperature tolerance as appeared in chlorophyll a fluorescence response. They showed a higher percentage increase in MDA levels, as compared to TTL-6, CRA-6017 and SMP-1, which showed low temperature susceptibility. But the reduction in the level of MDA by 4:30 p.m. (recovery) was faster in the clones TTL-1, TTL-4 and UPASI-9, as compared to TTL-6, SM/OM/54 and SMP-1. The result indicates that in TTL-6, SMP-1 and SM/ OM/54, the toxic oxygen species scavenging mechanisms may be less functional as compared to other clones. The percentage increase of proline and carotenoids was higher in clones TTL-1, TTL-4 and UPASI-9 as compared to TTL-6, CRA-6017 and SMP-1.