Uptake,transport and assimilation of 15N-nitrate and 15N-ammonium in tea (Camellia sinensis L.) plants

¹⁵N studies were conducted using hydroponically grown tea (Camellia sinensis L.) plants to clarify the characteristics of uptake, transport and assimilation of nitrate and ammonium. From the culture solution containing 50 mg L^-1 N0₃-N and 50 mg L^-1 NH.-N, the uptake of NH₃-N after 24 h was twice as high as that of NO₃-N, while the uptake of N0₃-N from the culture solution containing 90 mg N0₃-N and 10 mg NH₃-N was twice that of NH₄-N. The presence of 0.4 mM Al had no significant effect on the N0₃-N and NH₄-N uptake from the culture solutions containing 50 mg L^-1 N0₃-N and 50 mg L^-1 NH₄-N, 90 mg L^-1 N0₃-N and 10 mg L^-1 NH₄-N or 99 mg L^-1 N0₃-N and 1 mg L^-1 NH₄-N. Transport of N0₃-derived N to young leaves was much more rapid than that of NH₄-derived NO₃ and NH₄-derived N was largely retained in the roots and lower stem. Young and mature shoots separated from the roots absorbed more N0₃-N than intact plants. Nitrate assimilation occurred in both, roots and young as well as mature leaves. Internal cycling of N0₃-derived Nand NH₄-derived N from one root part to another part was not appreciable after 28 h, suggesting that a longer of time is required for cycling in woody plants.     

Stimulatory effect of aluminum on the growth of tea pollen tubes

It is well known that when inorganic elements such as boron (B), calcium, cobalt, and manganese are added to media in combination or alone, they stimulate the germination and/or tube growth of various kinds of pollen (Schmücker 1933; Loo and Hwang 1944; Yamada 1958; Brewbaker and Kwack 1963). Among these elements, the stimulatory effect ofB is the most effective and generally recognized. Aluminum (Al) belongs along with B to Group IIIa in the periodic table, and Fähnrich and Ultlich (1964) reported that Al inhibited the germination and growth of the pollen tube of Petunia hybrida and Antirrhinum majus. However, stimulatory effects of Al on pollen germination or pollen tube growth had not been previously reported.     

Assessment of aluminum sensitivity of maize cultivars using roots of intact plants and excised root tips

Maize cultivars (Zea mays L.) were evaluated for their aluminum (Al) sensitivity using intact plants and excised root tips exposed to 25 μM Al in nutrient solution of low ionic strength and pH 4.3. Aluminum supply increased callose formation and Al concentrations in root tips of intact plants as well as in excised root tips. Using intact plants, differences in Al sensitivity among cultivars could be characterized by Al‐induced callose formation, Al‐induced inhibition of root elongation, as well as Al contents in root tips as parameters. Significant correlations between Al‐induced callose formation and Al contents in root tips (r² = 0.64**) and inhibition of root elongation (r² = 0.80***) were found. Excised root tips did not show a significant Al‐induced inhibition of root elongation. While average Al‐induced callose formation was similar for root tips of intact plants and excised root tips, mean Al contents in excised root tips were up to 1.5‐fold higher than in root tips of intact plants after 24 h of Al treatment. Aluminum‐induced callose formation as found in excised root tips did neither correspond to Al‐induced callose formation nor to inhibition of root elongation of intact plants. The addition of 10 mM glucose to the incubation medium led to a significant increase in the elongation of excised root tips and a 2‐3‐fold increase in Al‐induced callose formation. Staining with triphenyl‐tetrazolium‐chloride (TTC) revealed increased viability of these root segments. However, these effects of glucose supply did not improve the characterization of the cultivars for Al resistance. The results presented suggest that Al exclusion mechanisms expressed in root tips of intact plants might be non‐operational in excised root tips. Therefore, the characterization of maize germplasm for Al resistance using excised root tips appears not to be reliable.     

Growth Characteristics of Tea Plants and Tea Fields in Japan

In 12th century, the Buddhist priest Eisai brought tea ( Camellia sinensis L.) seeds to Japan from China and now tea plants are cultivated all over Japan except in the Hokkaido and Tohoku districts. The quality (reflected in the price) of Japanese green tea is affected by the nitrogen content. Consequently in tea fields, for last three decades large amounts of fertilizer have been applied to produce high quality tea. As a result, problems such as acidification of soil have been caused. It is also known that the growth of tea plants is stimulated by the addition of aluminum (Al) under acidic conditions. In this keynote address, some problems caused by excess applications of fertilizer in tea fields and the growth characteristics of tea plants related to Al are presented.     

Effect of alternative anions (Cl– vs. SO$ _4^{2-} $) on concentrations of free amino acids in young tea plants

The quality of green tea is highly dependent on the concentration of free amino acids, whose profile is dominated by the unique amino acid theanine (N⁵‐ethyl‐glutamine). A high quality is associated with a high amino acid–to–catechin ratio, but previous results indicated that excessive chloride (Cl^–) supply is detrimental for amino acid accumulation. Several experiments were conducted to investigate the effect of chloride on growth and concentrations of free amino acids in young tea plants. Soil‐grown tea plants supplied with different levels of potassium (K) as K₂SO₄ or KCl exhibited increased concentrations of free amino acid in young shoots only when supplied with K₂SO₄, and the negative effect of KCl supply was mainly due to a reduced concentration of theanine. Concentrations of other nutrients in plant tissues were not influenced. The uptake of Cl^– and its interaction with nitrogen (N) uptake were further investigated in a second experiment, in which soil‐cultivated tea plants were supplied with varying amounts of Cl^–. Chloride application reduced yield of young shoots, and severity of leaf damage was related to the concentration of Cl^– in leaves. Nitrogen uptake was reduced by Cl^– addition. To verify whether the decrease of free amino acids was simply a result of inhibited NO assimilation, a third experiment was conducted, in which tea plants were NH ‐fed in the absence or presence (equivalent to the NH concentration) of Cl^–. Again, concentrations of theanine and total free amino acids in young shoots were reduced by Cl^– supply, but changes of the free–amino acid pool did not contribute to the maintenance of charge balance. However, concentration of theanine in roots, where it is synthesized, was not influenced by Cl^–. Total N concentrations of roots and mature leaves, uptake rate of NH , and activity of glutamine synthetase in fibrous roots and young leaves were all unaffected by Cl^– as well. It is suggested that translocation of theanine from root to shoot and its catabolism in young shoots might be influenced by Cl^–.     

永兴县发展冰糖橙产业的土地适宜性评价

为促进永兴县冰糖橙产业的持续快速发展,采用层次分析法、德尔斐法以及GIS空间叠加方法,对永兴县发展冰糖橙产业的土地适宜性进行了评价。结果表明:永兴县共有冰糖橙种植高度适宜性土地1 426.85hm2,适宜性土地28 858.43hm2,勉强适宜性土地406.57hm2,不适宜性土地4 639.71hm2,适宜性土地面积占土地总面积的85.72%,整体隶属度值为0.84,位于二级水平。基于土地评价结果,形成了永兴县冰糖橙适宜性评价体系,并提出了永兴县冰糖橙产业发展的建议。     

铝对花生根系膜脂过氧化和保护酶活性的影响

研究了铝对耐铝性不同的两个花生品种根系膜脂过氧化和保护酶活性的影响,结果表明,铝胁迫下,桂花21根系的超氧化物歧化酶(SOD)活性变化不大,而桂花23的SOD活性呈下降趋势;两个品种的过氧化物酶(POD)活性呈上升趋势,桂花21上升的幅度大于桂花23的;桂花21根系的过氧化氢酶(CAT)活性先升高后下降,桂花23的则下降;两个品种的脯氨酸和超氧阴离子自由基(O2-.)的含量均增加,但与对照相比,桂花21的脯氨酸升高的量大于桂花23的,桂花21 O2-.升高的量小于桂花23的;桂花23的丙二醛含量显著增加,而桂花21没有明显变化;两个品种根系电解质渗漏率均升高,桂花23上升的量大于桂花21的。     

Aluminium-induced callose formation in root tips of Norway spruce (Picea abies (L.) Karst.)

The objective of this study was to examine whether aluminium (Al) induces callose formation in roots of Norway spruce as it does in soybean. Spruce seedlings were grown in Al-free nutrient solution under controlled conditions in a growth chamber at pH 3.8. After 21 days 170 μM Al was added or not (controls) to the complete nutrient solution (molar Ca/Al ratio: 0.75). Callose could be detected in outer root-tip cells of Al-treated plants within 3 h, using fluorescence microscopy after staining with aniline blue. Prolonged Al treatment up to 24 h increased both the density of the callose deposits and the number of affected cell layers. Control plants showed no comparable callose deposits. Ultrastructural examinations showed cell-wall appositions in Al-treated root cells but not in controls. The possible implications of Al-induced callose formation for nutrient and water uptake by roots are discussed.     

CO2浓度升高对茶树光合速率影响的初步研究

通过人工调控CO2 浓度,对不同光环境下茶树(Camelliasinensis)的光合速率进行了研究。在气温34 0℃以上、光合有效辐射强度80 0 μmol/ (m2 s)左右的高温、强光环境中,提高大气CO2 浓度可缓解高温对光合速率的抑制作用,CO2 浓度提高到5 5 0 μl/l可使抑制茶树光合作用的气温提高约1～2℃,70 0 μl/l则可使抑制气温提高3～4℃。在强光下CO2 对茶树有较强的“气肥”作用,在光合有效辐射强度为4 0 0 μmol/ (m2 s)左右的弱光环境中CO2浓度提高也可提高光合速率水平。弱光环境中气温对光合作用的影响较强,茶树最适宜生长温度为2 9 0～30 0℃,气温超过30 0℃则温度开始对光合作用产生抑制,在气温达34 0℃以上高温时,茶树表现为净呼吸     

Tea plant (Camellia sinensis L.) roots secrete oxalic acid and caffeine into medium containing aluminum

We examined the response of the tea plant (Camellia sinensis L.) to aluminum (Al) exposure under sterile conditions, focusing specifically on the secretion of low molecular mass organic compounds from roots. After germination in agar medium, tea seedlings together with medium were placed on agar containing 0.4?mM Al with 0.2% hematoxyline (hematoxylin-Al medium). The purple color of the hematoxylin-Al medium was observed to fade gradually, until none of the color remained 6 days later. The tea seedlings were then treated with simple calcium solution (0.2?mM, at pH 4.2) containing AlCl₃, which ranged in concentration from 0 to 0.8?mM, for 24?hrs. The amount of oxalate secreted into the medium increased as the external Al concentration increased, while the concentrations of malate and citrate in the medium remained unchanged. Oxalate secretion started within 30?min after Al exposure and increased linearly thereafter. The findings demonstrated that oxalate was a key compound in the Al-tolerance mechanism employed by the tea plant, which detoxifies Al³⁺ externally in the rhizosphere. In addition to oxalate, caffeine was also secreted by tea roots in response to Al exposure. It is possible that caffeine excretion from the roots of tea plants may stimulate root growth through the inhibition of callose deposition in root tips.     

Effects of ferrous sulfate with sulfuric acid on tea (Camellia sinensis (L.) O. Kuntz) soil environment

In brown tea soil, the effects of ferrous sulfate with sulfuric acid on soil environment and tea plant (Camellia sinensis (L.) O. Kuntz) growth were examined by enzyme activity, microbe quantity, nutrients in rhizosphere soil, and the nutrients and dry matter weight in the tea plant. We found that the pH value in the brown soil was significantly decreased by ferrous sulfate with sulfuric acid treatments, compared with the value in unfertilized control (CK) or sulfuric acid treatments. The acid phosphatase activities of 3 g kg^–1 ferrous sulfate with 0.05 mL L^–1 sulfuric acid were increased by 210.53%. Interestingly, the quantities of fungi were increased by 19.76% and the bacteria and actinomyces were decreased in the CK treatment. Specifically, the nitrogen and magnesium in rhizosphere soil and leaves were significantly increased by the low concentrations of ferrous sulfate with 0.05 mL L^–1 sulfuric acid, whereas the dry matter weight under 3 g kg^–1 ferrous sulfate with 0.05 mL L^–1 sulfuric acid was increased by 36.22% in the CK treatment. Thus, we concluded that ferrous sulfate with sulfuric acid affected not only the soil environment but also the growth of tea plants. Our results suggested that the concentration of ferrous sulfate at 3 g kg^–1 soil diluted in 0.05 mL L^–1 sulfuric acid could be considered a good soil conditioner to make a suitable soil environment for tea production.     

茶树脱水素基因(CsDHN)的克隆及表达分析

脱水素(dehydrins,DHNs)是植物胚胎发育后期丰富蛋白(late embriogenesis abundant protein,LEA)中的一类,与低温、干旱以及盐胁迫等多种逆境反应相关.本研究采用cDNA末端快速扩增(rapidamplification of cDNA ends,RACE)技术从茶树(Camellia sinensis)中获得一种茶树脱水素基因CsDHN (GenBank登录号:FJ436978),序列分析显示,该基因cDNA全长960 bp,编码201个氨基酸,推测编码蛋白质分子量约为21kD,等电点为8.3,属于Y3SK2型脱水素.CsDHN基因有2个外显子和1个内含子.生物信息学预测显示,该脱水素蛋白亲水性强、无信号肽和跨膜区,亚细胞定位于细胞质中.表达特性分析表明,CsDHN在逆境及脱落酸(abscisic acid,ABA)诱导下可上调其转录水平,4℃低温处理下表达量可提高4.2倍,而脱水处理能提高93.4倍,高盐处理能提高17.8倍,CsDHN对脱水、高盐胁迫的响应程度优于低温胁迫.CsDHN在茶树各组织器官中都有表达,其中种子中表达量最高,为叶片的11.2倍.研究表明脱水素基因CsDHN可能在茶树防御非生物逆境胁迫和参与茶树种子成熟脱水的活力保护过程起一定作用,为了解茶树抗逆分子机制提供了一定的理论基础.     

Microsatellite variability within and among local landrace populations of tea, Camellia sinensis (L.) O. Kuntze, in Kyoto, Japan

Genetic variability within and among eight landrace populations of tea (Camellia sinensis (L.) O. Kuntze) located in southern Kyoto, Japan, was surveyed with six microsatellite markers. The average number of alleles per locus was 3.83 to 4.67 for landrace populations, whereas the corresponding value among modern cultivars and breeding lines was 6.63. Expected heterozygosity values averaged over loci within landrace populations ranged from 0.498 to 0.723. A similar level of variation, 0.682, was observed for cultivars and breeding lines. High fixation index values (0.177–0.417) for each population are consistent with biparental inbreeding within the population. Genetic differentiation among local populations was extremely low with F _ST = 0.062, although AMOVA revealed significant differentiation among landrace populations. We propose that these populations share a common ancestral gene pool and that some degree of artificial selection within each population has been performed by local farmers. Neighbor-joining analysis revealed that genetic relationships among populations reflect geographical location of populations. This might result from more frequent genetic exchange by nearby farmers.     

Enhancement of callose production by a combination of aluminum and iron in suspension-cultured tobacco (Nicotiana tabacum) cells

In nutrient medium, aluminum (A1) enhances ferrous ion [Fe(II)] -mediated per oxidation of lipids, which results in the loss of the plasma membrane integrity and the accumulation of A1 in tobacco cells. Under these conditions, the mechanism of callose production and possible involvement of callose in the accumulation of Al were investigated. Callose production was enhanced by both Al and Fe(II), but not by A1 or Fe(II) alone, and the enhancement was inhibited by a lipophilic antioxidant, suggesting that the enhancement of callose production is caused by the A1-enhanced, Fe(II)-mediated peroxidation of lipids. The enhancement of callose production depended on the presence of external Ca²⁺ in the treatment medium. The activity of β-l,3-glucan synthase in the microsomes was increased several times by the addition of Ca²⁺ in the assay medium, although the activity in the microsomes was reduced by the treatment of cells with Al and Fe(II) together. Therefore, it is likely that callose production is enhanced by exogenous Ca²⁺ via the AI-enhanced, Fe(II)-mediated peroxidation of lipids. During the exposure of the cells to Al and Fe(II), callose production started and increased simultaneously with Al accumulation. However, the digestion of callose in the cell wall materials prepared from the A1-treated cells by laminarinase did not release A1, suggesting that callose is not involved in the binding or trapping of A1.     

茶树生长素抑制蛋白基因CsARP1的克隆与表达分析

从茶树休眠芽抑制消减杂交文库中分离得到生长素抑制蛋白基因的3＇-片段,以休眠芽为材料,利用RACE技术克隆了其cDNA全长,并利用荧光定量PCR研究了该基因在不同休眠阶段芽的相对表达量。结果从茶树休眠芽中获得一个全长为711bp的生长素抑制蛋白基因CsARP1（GenBank登录号为HQ225758）。该基因开放阅读框...     

Dynamics of nitrogen translocation from mature leaves to new shoots and related gene expression during spring shoots development in tea plants (Camellia sinensis L.)

The contribution of N remobilization is crucial for new shoots growth and quality formation during spring tea shoots development. However, the translocation mechanism of N from source leaves to sink young shoots is not well understood. In the present study, ¹⁵N urea was applied to mature tea leaves one week before bud break to track N remobilization in a field experiment. The dynamic changes in plant ¹⁵N abundance, contents of amino acids, and the expression levels of genes related to N metabolism and translocation were followed during the 18‐d development of new spring shoots until three expanding young leaves. The results showed that during the growth of new shoots the amount of ¹⁵N in the shoots increased, whereas the N_dff (N derived from ¹⁵N‐urea) in mature leaves decreased, showing that the foliar‐applied N in mature leaves was readily exported to new shoots. This process was found to be accompanied by decline of chlorophylls. In the mature leaves, expression CsATG18a and CsSAG12 involved in autophagy was dramatically induced (> 4‐fold) at approximately nine days after the bud breaking. The genes involved in the transformation of amino acids, including primarily CsGDH2, CsGDH4, CsGLT3, CsGS1;3, and CsASN2 were upregulated by > 3‐fold after bud breaking. The expression levels of CsATG8A, CsATG9, CsSAG12, CsGS1;1, CsGDH1, and CsAAP6 correlated negatively with the N_dff in mature leaves, but positively with ¹⁵N amount and total N amount in new shoots, suggesting these genes played important roles in N export from mature leaves. In the new shoots, the expression of most genes showed two defined peaks, one on six days and one on 12 days after bud breaking. The expression of CsGS2, CsASN3, CsGLT1, and CsAAP4 positively correlated with the ¹⁵N amount and total N amount in new shoots. These genes might be involved in the transport and re‐assimilation of N from mature leaves. The overall results demonstrated that the translocation of ¹⁵N from mature leaves to new spring shoots was regulated by the genes involved in autophagy, protein degradation, amino acid transformation and transport.     

铝对小麦根尖细胞壁过氧化物酶活性和过氧化氢含量的影响

以2个小麦基因型鉴-864(耐性)和扬麦5号(敏感)为材料,采用溶液培养方法研究了铝胁迫下小麦根系伸长、根尖铝含量、根尖细胞壁过氧化物酶活性和H2O2含量的变化。结果表明,随着铝浓度的提高,小麦根系伸长受铝抑制程度加剧,根尖铝含量也明显升高;但敏感基因型根尖铝含量较高,根系伸长受抑程度更为明显。在铝胁迫下,2个小麦基因型根尖可溶态愈创木酚过氧化物酶(GPX)和松柏醇过氧化物酶(CAPX)活性没有显著变化,细胞壁离子键结合态GPX和CAPX的活性则随着铝浓度的提高而显著升高,H2O2含量也呈现类似的趋势;而敏感基因型过氧化物酶活性升高和H2O2积累更为明显。因此,铝胁迫下,小麦敏感基因型根尖细胞壁离子键结合态GPX和CAPX活性升高而引起H2O2积累,促进根系木质化和细胞壁氧化交联,导致根细胞壁刚性提高而伸展能力降低,是其根系伸长受到严重抑制的原因。     

The coupling effect of water and fertilizer on the growth of tea plants [Camellia sinensis (L.) O. Kuntz]

Water and fertilizer are the two main factors promoting the rapid growth of tea plants (Camellia sinensis). Using two-year-old rooted cutting of Wuniuzao (Camellia sinensis) as experimental materials, “311-B” D-saturation optimization design was carried out using 5, 4, and 4 levels of irrigation regimes, fertilizer nitrogen, and fertilizer phosphorus, with six replications. The raw date obtained was used of quadratic regression fitting for the plant height, new-tip length and dry matter weight. The coupling effects of these three factors [nitrogen (N0, water, and phosphorus (P)] on dry matter weight were significant. The effect order of single factor on dry matter weight was N>water>P, while the interactive effect order on the dry matter weight was N and water>P and water>N and P. An optimal combination with 0.485 g·pot^?1 N, 0.274 g·pot^?1 P, and once every 4-5 days of watering was proposed for the highest tea dry matter yield.     

Aluminum inhibits apoplastic flow of high–molecular weight solutes in root apices of Zea mays L.

Using an aluminum (Al)‐sensitive maize cultivar, we investigated the influence of Al on the apoplastic solute bypass flow and its relationship with Al‐induced (1 h, 50 μM) callose formation and root growth. We selected the fluorescent probes 8‐hydroxypyrene‐1,3,6‐trisulfonic acid, trisodium salt (MW 524) (HPTS) and dextran‐Texas Red (TR) conjugates (MW 3,000, 10,000, and 40,000) to monitor their apoplastic transport. Confocal laser–scanning microscopy (CLSM) analysis and spectrofluorometric quantification showed Al‐induced callose formation in peripheral root cells within 1 h. Pretreatment of plants with the callose synthesis inhibitor 2‐deoxy‐D‐glucose (DDG) reduced the callose formation by half. Uptake experiments with both HPTS and dextrans showed uniform dye distribution in control root apices. After Al treatment for 1 or 2 h, which inhibited root growth by 32% or 50%, respectively, the dyes accumulated in the epidermal and outer cortical cell layers, especially in the 1–2 mm apical root zone. Al treatment reduced the export of the dyes out of the apical 1 cm treatment zone. This was due to strong sorption of HPTS but not of dextrans by Al‐loaded cell walls. Aluminum treatment reduced loading into the xylem sap particularly of higher–molecular weight dextrans. Pretreatment of roots with DDG and presence of 50 mM mannitol during the Al treatment partially forestalled the inhibitory effect of Al on the dye transport, but only slightly reduced the Al‐induced growth inhibition. Exudation experiments revealed that xylem water flow remained unaffected by the Al treatment of the root tips. The results with dextran suggest that Al binding in cell walls of the root apex inhibits apoplastic bypass flow of higher–molecular weight solutes, which might contribute to Al‐induced inhibition of root growth.