甘薯块根膨大过程中ATP酶活性、ATP和ABA含量的变化

研究选用鲁薯7号和徐薯18号为材料,对甘薯块根膨大速率变化动态及其块根中可溶性碳水化合物含量、ATP含量、ATP酶活性和脱落酸（ABA）含量的变化进行了研究分析。结果表明：（1）块根膨大速率变化动态呈一双峰曲线,第一个高峰出现在栽秧后50－70d,第二个高峰出现在栽秧后120－165d;（2）块根膨大高峰期,块根中可溶性碳水化合物含量较高,ATP含量则较低;（3）块根中ATP酶活性和ABA含量变化动态与块根膨大速率变化动态相似。讨论了ATP酶和ABA在块根膨大过程中的可能作用。     

Increase in mitochondrial activity in diseased sweet potato root tissue

When infected with Ceratocystis fimbriata, sweet potato roottissue showed an increase in respiration concomitant with anincrement in mitochondrial activities, which was not in parallelwith the increase in mitochondrial numbers. ¹ This paper constitutes part 100 of the phytopathological chemistryof sweet potato with black rot and injury. ² Present address: Department of Biochemistry, Faculty of NaturalSciences, Komensky University, Bratislava, Czechoslovakia. (Received June 21, 1972; )     

Involvement of nitrogen in storage root growth and related gene expression in sweet potato (Ipomoea batatas)

• Nitrogen (N) could affect storage root growth and development of sweet potato. To manage external N concentration fluctuations, plants have developed a wide range of strategies, such as growth changes and gene expression.
• Five sweet potato cultivars were used to analyse the functions of N in regulating storage root growth. Growth responses and physiological indicators were measured to determine the physiological changes regulated by different N concentrations. Expression profiles of related genes were analysed via microarray hybridization data and qRT‐PCR analysis to reveal the molecular mechanisms of storage root growth regulated by different N concentrations.
• The growth responses and physiological indicators of the five cultivars were changed by N concentration. The root fresh weight of two of the sweet potato cultivars, SS19 and GS87, was higher under low N concentrations compared with the other cultivars. SS19 and GS87 were found to be having greater tolerance to low N concentration. The expression of N metabolism and storage root growth related genes was regulated by N concentration in sweet potato.
• These results reveal that N significantly regulated storage root growth. SS19 and GS87 were more tolerant to low N concentration and produced greater storage root yield (at 30 days). Furthermore, several N response genes were involved in both N metabolism and storage root growth.

     

Analysis of genes developmentally regulated during storage root formation of sweet potato

To identify the genes involved in storage root formation of sweet potato (Ipomoea batatas), we performed a simplified differential display analysis on adventitious roots at different developmental stages of the storage root. The expression patterns were confirmed by semiquantitative RT-PCR analyses. As a result, 10 genes were identified as being developmentally regulated and were named SRF1-SRF10. The expression of SRF1, SRF2, SRF3, SRF5, SRF6, SRF7, and SRF9 increased during storage root formation, whereas the expression of SRF4, SRF8, and SRF10 decreased. For further characterization, a full-length cDNA of SRF6 was isolated from the cDNA library of the storage root. SRF6 encoded a receptor-like kinase (RLK), which was structurally similar to the leucine-rich repeat (LRR) II RLK family of Arabidopsis thaliana. RNA gel blot analysis showed that the mRNA of SRF6 was most abundantly expressed in the storage roots, although a certain amount of expression was also observed in other vegetative organs. Tissue print mRNA blot analysis of the storage root showed that the mRNA of SRF6 was localized around the primary cambium and meristems in the xylem, which consist of actively dividing cells and cause the thickening of the storage root.     

Changes in glutamate dehydrogenase activity in cut-injured sweet potato root tissue

Changes in glutamate dehydrogenase activity in sweet potatoroot tissue in response to slicing were investigated with mitochondrialand supernatant fractions. Results indicated that activity inmitochondrial fraction had decreased, whereas activity in supernatantfraction had increased, 12 hr after slicing. The increase inactivity in the supernatant fraction may be related to the regenerationof phenylalanine, a precursor of polyphenols. ¹This paper constitutes Part 104 of the Phytopathological Chemistryof Sweet Potato with Black Rot and Injury. ²Present address: Laboratory of Biochemistry, Faculty of Agriculture,University of Tokyo, Bunkyoku, Tokyo 113, Japan. (Received August 15, 1972; )     

Establishment and characteristics of an anthocyanin-producing cell line from sweet potato storage root

 Anthocyanin pigments accumulated in a cell line derived from storage-root explants of sweet potato (Ipomoea batatas L.) cv 'Ayamurasaki'. Somatic pro-embryos were induced on the explants cultured on Murashige and Skoog medium supplemented with 1 mg/l 2,4-D. The pro-embryo structures produced callus when transferred to MS medium with 0.5 mg/l 2,4-D. A cell line was isolated from this callus which accumulated anthocyanin pigment. The color value of the pigment extracted after 27 days of culture in MS medium with 2 mg/l 2,4-D was 8.2, which was very close to that of a pigment extracted from roots, which was 8.9. Most of the pigments from the cell extract were hydrophilic and appeared on the ODS-column HPLC with a lower retention time than the main anthocyanins of the root tissues. The majority of the pigments were identical with the root anthocyanins. Cell line-specific anthocyanins were detected. Received: 8 January 1999 / Revision received: 2 March 1999 / Accepted: 30 June 1999     

甘薯蔗糖转运蛋白的功能分析

甘薯(Ipomoea batatas)是重要的粮食和工业加工原料作物。蔗糖是植物体内碳水化合物长距离转运的主要形式,蔗糖转运蛋白(sucrose transporter,SUT)在植物的生长代谢中调控蔗糖的跨膜运输和分配,在韧皮部介导的源-库蔗糖运输和为库组织供应蔗糖的生理活动中起关键作用。本研究根据不同淀粉性状甘薯块根中差异表达的2个SUT基因转录本,进行cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)克隆,获得IbSUT62788和IbSUT81616的全长cDNA序列;通过系统发育分析明确其分类;通过在本氏烟草(Nicotiana benthamiana)中瞬时表达明确其亚细胞定位;通过酵母功能互补系统鉴定IbSUT62788和IbSUT81616是否具有吸收、转运蔗糖和己糖的能力。通过实时荧光定量PCR(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)分析IbSU62788和IbSUT81616在甘薯各器官中的表达特征;通过蘸花法得到外源表达IbSUT62788和IbSUT81616基因的拟南芥(Arabidopsis thaliana)植株,比较与野生型拟南芥的淀粉和糖含量的差异。结果表明,IbSUT62788和IbSUT81616分别编码505个和521个氨基酸的SUT蛋白,均属于SUT1亚家族。IbSUT62788和IbSUT81616均定位于细胞膜,在酵母系统中具有转运蔗糖、葡萄糖和果糖的能力。此外,IbSUT62788还具有转运甘露糖的能力。IbSUT62788在甘薯叶片、侧枝和茎中的表达量更高,IbSUT81616在侧枝、茎和块根中表达量更高。IbSUT62788和IbSUT81616在拟南芥中异源表达后,植株可以正常生长,但生物量增加。IbSUT62788的异源表达增加了拟南芥植株叶片可溶性糖含量、叶片大小和种子千粒重;IbSUT81616的异源表达增加了拟南芥植株叶片、根尖的淀粉积累量和种子千粒重,但减少了可溶性糖含量。本研究结果表明,IbSUT62788和IbSUT81616可能是调控甘薯蔗糖和糖含量性状的重要基因,在细胞膜上进行着蔗糖的跨膜运输、蔗糖进出库组织、韧皮部蔗糖的运输与卸载等生理功能,在拟南芥中异源表达造成的性状改变说明其在提高其他植物或作物产量中的应用潜力。本研究为揭示甘薯淀粉和糖代谢及重要品质性状形成机制提供了重要信息。     

Regulated expression of ADPglucose pyrophosphorylase and chalcone synthase during root development in sweet potato

The phase transition in sweet potato root during tuber differentiationis a complex developmental process that involves changes in gene expression andmorphogenesis. Among the three kinds of root in sweet potato (white fibrousroot, thick-pigmented root and lateral root), ADP-glucose pyrophosphorylase(AGPase) and chalcone synthases (CHS) are expressed only in thick-pigmentedroots after 3 weeks, and this also depends on the developmental stage. Sinceexposing roots to the light or culturing under hydroponic conditions inhibitstuber formation in sweet potato, the expression of AGPase and CHS was studiedunder light and dark conditions. AGPase and CHS expression in sweet potatorootswas suppressed very sensitively by light or water stress, similar to rootdevelopmental patterns. Based on an analysis of AGPase and CHS expression indifferent kinds of root tissues and in different developmental stages, thesegenes were shown to be closely associated with the differentiation ofthickeningpigmented roots.     

Biosynthesis of ethylene in sweet potato root tissue

The biosynthetic pathway of ethylene in freshly cut and blackrot-diseased tissues of sweet potato roots was investigated.Glucose-U-¹⁴C administration gave labeled ethylene in both freshand diseased tissues, but at the early stage of infection, therewas ethylene production which was not derived from the fed ¹⁴C-glucose.Acetate-1-¹⁴C and acetate-2-¹⁴C were equally incorporated intoethylene produced from fresh tissue, but acetate-2-¹⁴C was preferentiallyincorporated into ethylene from diseased tissue. Pyruvate-3-¹⁴Cwas more efficient as a precursor than was acetate or glucosein fresh tissue, while its efficiency was the same as that ofacetate in diseased tissue. Monofluoroacetate promoted pyruvate-3-¹⁴Cincorporation in fresh tissue but inhibited incorporation indiseased tissue. We concluded that the TCA cycle is involvedin the case of diseased tissue but not in fresh tissue; thus,showing different pathways for ethylene production in each tissue.In addition, in diseased tissue, ethylene is assumed to be producedfrom some cellular component(s), not easily synthesized fromglucose through fungus infection, but is degraded as soon asinfection commences. ¹This paper constitutes Part 85 of the Phytopathological Chemistryof Sweet Potato with Black Rot and Injury ²Present address: The Institute for Biochemical Regulation,Faculty of Agriculture, Nagoya University, Chikusa, Nagoya 464,Japan (Received April 20, 1970; )     

Root and shoot initiation by leaf,stem, and storage root explants of sweet potato

Explants from stem, leaf, and storage root tissue of sweet potato (Ipomoea batatas L.) cv. Jewel, were placed on media conaining 0.1, 1.0, and 10 mg/1NAA with 0.1, 1.0, or 10 mg/1BA in a factorial experiment. Some callus formed in every treatment, but the best callus growth was on media containing 1.0 mg/1NAA and 10 mg/1BA. Roots formed over a range of treatments but were most prolific on the medium containing 1.0 mg/1NAA and 0.1 mg/1BA. Some de novo formed roots subsequently produced shoot buds in culture. Shoot formation increased the longer the original explants remained in culture without subculture. Roots could be subcultured indefinitely on agar solidified medium, but shoot regeneration did not occur after two subcultures. Shoot formation was greatest when the roots were subcultured on medium containing 1.0 mg/NAA and 0.1 mg/1BA. The cultivar Caromex followed the same regeneration pathway, but the number of shoots formed was considerably less. Regeneration in both Jewel and Caromex explants was enhanced by light.Paper No. 8292 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC. The use of trade names in this publication does not imply endorsement by the North Carolina Agricultural Research Service of products named, nor criticism of similar ones not mentioned.This work was done as a partial requirement for the M.S. degree at North Carolina State University.     

Sucrose metabolism in cold-stored potato tubers with decreased expression of sucrose phosphate synthase

Transfer of potato tubers to low temperature leads after 2–4 d to a stimulation of sucrose synthesis, a decline of hexose-phosphates and a change in the kinetic properties, and the appearance of a new form of sucrose phosphate synthase (SPS). Antisense and co-suppression transformants with a 70–80% reduction in SPS expression have been used to analyse the contribution of SPS to the control of cold sweetening. The rate of sucrose synthesis in cold-stored tubers was investigated by measuring the accumulation of sugars, by injecting labelled glucose of high specific activity into intact tubers, and by providing 50 mol m^–3 labelled glucose to fresh tuber slices from cold-stored tubers. A 70–80% decrease of SPS expression resulted in a reproducible but non-proportional (10–40%) decrease of soluble sugars in cold-stored tubers, and a non-proportional (about 25%) inhibition of label incorporation into sucrose, increased labelling of respiratory intermediates and carbon dioxide, and increased labelling of glucans. The maximum activity of SPS is 50-fold higher than the net rate of sugar accumulation in wild-type tubers, and decreased expression of SPS in the transformants was partly compensated for increased levels of hexose-phosphates. It is concluded that SPS expression per se does not control sugar synthesis. Rather, a comparison of the in vitro properties of SPS with the estimated in vivo concentrations of effectors shows that SPS is strongly substrate limited in vivo . Alterations in the kinetic properties of SPS, such as occur in response to low temperature, will provide a more effective way to stimulate sucrose synthesis than changes of SPS expression.     

Gene expression profiles of the one-carbon metabolism pathway

One-carbon metabolism plays a critical role in both DNA methylation and DNA synthesis. Accumulating evidence has shown that interruptions of this pathway are associated with many disease outcomes including cardiovascular diseases and cancers. Mechanistic studies have been performed on genetic polymorphisms involved in one-carbon metabolism. However, expression profiles of these inter-related genes are not well-known. In this study, we examined the gene expression profiles of 11 one-carbon metabolizing genes by quantifying the mRNA level of the lymphocyte among 54 healthy individuals and explored the correlations of these genes. We found these genes were expressed in lymphocytes at moderate levels and showed significant inter-person variations, We also applied principle component analysis to explore potential patterns of expression. The components identified by the program agreed with existing knowledge about one-carbon metabolism. This study helps us better understand the biological functions of one-carbon metabolism.     

Structural relationship among the members of a multigene family coding for the sweet potato tuberous root storage protein

Sporamin, the major soluble protein of the sweet potato tuberous root, is coded for by a multigene family. Fourty-nine essentially full-length sporamin cDNAs isolated from tuberous root cDNA library have been classified by cross hybridization, restriction endonuclease cleavage pattern and ribonuclease cleavage mapping. All the cDNAs fall into one of the two distinct homology groups, subfamilies A and B, which correspond to the polypeptide classes sporamin A and B, respectively. At least 5 different sequences are detected in both of the 22 sporamin A and 27 sporamin B cDNAs. Comparison of the nucleotide sequences of the coding region of three each of sporamin A and B subfamily members, four from cDNAs and two from genomic clones, indicates that intra-subfamily homologies (94 to 98%) are much higher than inter-subfamily homologies (82 to 84%), and there are deletions or insertions of one or two codons at three locations which characterize each subfamily. Large portions of base substitutions in the coding region accompany amino acid substitutions. In contrast to the coding region, most of the structural differences among the members in the 5 and 3 noncoding regions are deletions or insertions.     

Enhancing vacuolar sucrose cleavage within the developing potato tuber has only minor effects on metabolism

Modification of tuber carbohydrate metabolism by the tuber-specific expression of a yeast invertase targeted to the cytosol or apoplast has previously been demonstrated to have diverse effects on tuber growth and metabolism. In the current study, we generated plants exhibiting tuber-specific expression of the same enzyme targeted to the vacuole. Enzymatic analysis of the carbohydrate levels of the tuber revealed dramatic decreases in sucrose content coupled with large increases in the levels of glucose and hexose phosphates, but unaltered starch content in the transformants. Analysis of the key enzyme of glycolysis suggests that this pathway is down-regulated in the transformants. Despite these changes in metabolite pools and enzyme activity, few consistent changes could be observed in the estimated metabolic fluxes following incubation of isolated tuber discs in labelled glucose. The analysis of the relative levels of a wide range of metabolites using a gas chromatography-mass spectrometry (GC-MS)-based metabolite profiling method revealed large changes in the levels of fructose and decreases in a range of other sugars, but very few changes in the contents of organic and amino acids. This metabolic profile is remarkably consistent with that obtained following expression of the invertase in the apoplastic compartment, providing circumstantial evidence for the endocytotic trafficking of sugars within potato tuber parenchyma. Finally, the results of this study are compared with those from other plant species and the relative roles of the vacuolar isoform of the enzyme are contrasted.     

Effects of prolonged restriction in water supply on photosynthesis, shoot development and storage root yield in sweet potato

Besides the paucity of information on the effects of drought stress on photosynthesis and yield in sweet potato [ Ipomoea batatas (L.) Lam.], available reports are also contradictory. The aim of this study was to shed light on the effects of long-term restricted water supply on shoot development, photosynthesis and storage root yield in field-grown sweet potato. Experiments were conducted under a rainout shelter where effects of restricted water supply were assessed in two varieties (Resisto and A15). Large decreases in stomatal conductance occurred in both varieties after 5 weeks of treatment. However, continued measurements revealed a large varietal difference in persistence of this response and effects on CO₂ assimilation. Although restricted water supply decreased leaf relative water content similarly in both varieties, the negative effects on stomatal conductance disappeared with time in A15 (indicating high drought acclimation capacity) but not in Resisto, thus leading to inhibition of CO₂ assimilation in Resisto. Chlorophyll a fluorescence measurements, and the relationship between stomatal conductance, intercellular CO₂ concentration and CO₂ assimilation rate, indicated that drought stress inhibited photosynthesis primarily through stomatal closure. Although yield loss was considerably larger in Resisto, it was also reduced by up to 60% in A15, even though photosynthesis, expressed on a leaf area basis, was not inhibited in this variety. In A15 yield loss appears to be closely associated with decreased aboveground biomass accumulation, whereas in Resisto, combined effects on biomass accumulation and photosynthesis per unit leaf area are indicated, suggesting that research aimed at improving drought tolerance in sweet potato should consider both these factors.     

Production of ethylene by injured sweet potato root tissue

Ethylene production by sweet potato root tissue was examinedwith special emphasis on tissue injury. The root tissue producedethylene in response to cut injury. Increasing the cut surfacearea increased ethylene production, and the amount was proportionalto logarithm of the surface area. Tissue discs washed with waterbefore incubation produced less ethylene than unwashed discs. When the tissue was treated with chemicals that might destroythe cells, ethylene production remarkably increased. Monoiodoacetamide,trichloroacetic acid and sodium ethylmercurithiosalicylate wereparticularly effective in inducing ethylene production. Here,again, ethylene production was related to the degree of injury.Treatment of the tissue with increasing concentrations of thesechemicals resulted in increasing ethylene production, but concentrationsover a certain limit rather decreased the ethylene production.This may be due to the rapid destruction of the whole tissueused before ethylene production commenced. For thylene production,the presence of injured but still living cells was necessary. Relationship of the injury-induced ethylene production to metabolicactivation is discussed. ¹Part 67 of the Phytopathological Chemistry of Sweet Potatowith Black Rot and Injury. ²Fulbright grantee of 1967. Present address: Department of Biochemistry,University of Wisconsin, Madison, Wisconsin, U.S.A.