Optimal Nitrogen Content and Photosynthesis in Cauliflower (Brassica oleracea L. botrytis). Scaling up from a Leaf to the Whole Plant

A simple model of photosynthesis is described which is dependenton leaf area, organic nitrogen content and distribution withinthe canopy as well as on the light and temperature environments.The model is parameterized using a cauliflower crop as an example.The optimized protein-N profile within the canopy is calculatedwith respect to daily growth rate. By comparison with measuredprotein-N contents, the amount of super-optimal N-uptake, i.e.the N-uptake which does not increase productivity, is assessedfor two different nitrogen and light treatments. The amountof super-optimal N accumulated in cauliflower depends on N-supplyand can exceed 80 kg N ha^-1. Copyright 2000 Annals of BotanyCompany Brassica oleracea L. botrytis, cauliflower, nitrogen, photosynthesis, respiration, model, optimization     

Serine Transhydroxymethylase of Cauliflower (Brassica oleracea var. botrytis L.): Partial Purification and Properties

Serine transhydroxymethylase (EC 2.1.2.1) has been purified 46-fold from cauliflower (Brassica oleracea var. botrytis L.). The enzyme was completely dependent on the presence of tetrahydrofolic acid for the conversion of serine to glycine. The addition of pyridoxal phosphate gave a large increase in the reaction rate. A double pH optimum was observed with maxima at 7.5 and 9.5. The enzyme is specific for l-serine. The d-isomer is neither a substrate nor an inhibitor. The Michaelis constants for l-serine, tetrahydrofolic acid, and pyridoxal phosphate were 300 μm, 760 μm, and 24 μm, respectively. The addition of K⁺ also stimulated the reaction rate considerably. The effect was quite specific since all other metal ions tested either had very little: influence or were extremely inhibitory.     

Evaluation of Abiotic Stress Resistance in Mutated Populations of Cauliflower (Brassica oleracea var. botrytis)

An efficient in vitro screening method has been developed for cauliflower to create NEU and NMU induced mutant lines selected on hydroxyproline containing medium. Mutant lines and control plants were sub-cultured many times on maintenance medium and stored at 5°C for 2 years and then tested for salt and hydroxyproline resistance as in vitro and in vivo plants. In vitro shoot tips were also sub-cultured to media containing hydroxyproline and NaCl for 28 days and then assessed for their leaf proline content. Non-acclimated and acclimated in vivo plants were also assessed for resistance to freezing. Populations of control and selected lines were created by mass pollination and subsequently tested for their NaCl and frost resistance. Control plants had little or no NaCl or hydroxyproline resistance whilst selected plants showed varying degrees of resistance. In vitro and in vivo responses of selected lines were correlated. Leaf proline content was increased markedly in the mutant lines and the greatest proline contents occurred following NaCl stress with the most respondent line having 100-fold levels compared to the controls. Both non-acclimated and acclimated selected lines showed improved frost resistance over controls. Improvements in frost resistance were heritable but improvements in NaCl resistance were not. The results clearly demonstrated that NaCl, frost and hydroxyproline resistance were stable traits over repeated in vitro sub-cultures and prolonged low temperature storage. A complete range of mutants with single, double or triple resistance traits were produced. The level of resistance however was not necessarily correlated with the level of proline and some lines showed resistance without elevated proline. It is concluded that elevated proline is not essential for improved resistance to abiotic stress in cauliflower, but where it does occur it does improve resistance.     

Subcellular Distribution of O-Acetylserine(thiol)lyase in Cauliflower (Brassica oleracea L.) Inflorescence

The subcellular localization of O-acetyiserine(thiol)lyase (EC 4.2.99.8) in nongreen tissue from higher plants has been studied using purified proplastids, mitochondria, and protoplasts from cauliflower (Brassica oleracea L.) buds as a source of subcellular fractions. O-Acetylserine(thiol)lyase has been detected in both organelles (proplastids and mitochondria) and a cytosolic extract obtained by protoplast fractionation. We confirmed these observations, demonstrating that a form of the enzyme different in global charge and separated from others by anion-exchange chromatography corresponded to each subcellular location. Our observations are consistent with the need for cysteine biosynthesis in each subcellular compartment where the synthesis of proteins occurs.     

Effects of temperature and irradiance on vegetative growth of cauliflower (Brassica oleracea L. botrytis) and broccoli (Brassica oleracea L. italica)

Cauliflower (Brassica oleracea L. botrytis) and broccoli (Brassicaoleracea L. italica) plants were grown in large pots in growthchambers for a range of temperatures (mean air temperaturesfrom 7.0-25.3 C) and irradi-ances (from 9.3-50.8 mol m^–2d^–1 or 4.7-25.4 MJ m^–2 d^–1). The extinctioncoefficient for PAR decreased with plant size reaching a valueof 0.55 in cauliflower and 0.45 in broccoli at plant leaf areasof 0.235 m² and 0.227 m², respectively. The leaf area expansionrate was unaffected by irradiance when compared at identicalleaf surface temperatures. The response of expansion rate tosurface temperature was fitted to a broken stick model witha base temperature of –0.7C and an optimum temperatureof 21.0C. The radiation conversion coefficient increased withair temperature below 13.8C and remained constant above this.The estimated radiation conversion coefficient above 13.8Cand for a PPFD of 20 mol m^–2 d^–1 was 0.77 g mol^–1in cauliflower and 0.87 g mol^–1 in broccoli. The radiationconversion coefficient declined with increasing irradiance levelfrom a maximum of 1.89 g mol^–1 at near nil irradiancein cauliflower. Key words: Leaf area, dry matter, radiation use efficiency, extinction coefficient     

花椰菜BobACT基因的克隆及其作为内参基因的研究

实时荧光定量PCR技术是探索植物基因功能和调节机理的有效手段。选择合适的内参基因是获得实时荧光定量PCR准确性数据的必备条件。ACT基因高度保守且表达稳定,常作为内参基因被广泛应用。为了获得花椰菜ACT基因,以转录组测序和RT-PCR方法为手段克隆得到花椰菜肌动蛋白基因Actin。该基因等电点为5.395,理论分子量为41.77 kD;其cDNA开放阅读框长1134 bp,编码氨基酸377个,GenBank登录号为MG598643。Wolf Psort分析发现,BobActin蛋白亚细胞定位于细胞质基质中。Motif Scan分析显示,BobActin蛋白质的氨基酸序列4～377位为Actin保守结构域。进化分析表明,同源序列基因编码的蛋白质与同为十字花科的甘蓝、芜菁和油菜同源蛋白的相似性达到90%以上,具有高度的保守性。在此基础上,设计了1对荧光定量PCR引物,分析显示,该引物具有较高的特异性和扩增效率,在花椰菜根、茎、花、花球、叶片等不同组织和低温、高温、盐处理、干旱处理、ABA处理等胁迫处理下均能稳定表达,适合在花椰菜基因表达研究中作为内参基因,为开展花椰菜重要功能基因的挖掘、表达模式以及调控机理的研究提供参考。花椰菜在内参基因方面的研究还处于初步阶段,今后可继续克隆其他内参基因,丰富花椰菜的内参基因库,从而进一步提高花椰菜基因表达分析研究的稳定性、重复性和准确性。     

影响花椰菜农杆菌介导转化因素的研究

以花椰菜赛雪的带柄子叶为外植体,以MS为基本培养基,GUS基因为报告基因,分析了遗传转化过程中的影响因子,如预培养时间、农杆菌菌液浓度、侵染时间、共培养时间、乙酰丁香酮浓度、延迟筛选时间等对外植体瞬间表达和稳定表达的影响。结果显示,以花椰菜的带柄子叶为外植体,预培养2d,农杆菌菌液为OD6000.3～0.4,侵染8min,共培养2d,乙酰丁香酮浓度为100μmol/L,延迟筛选7d,卡那霉素筛选压为5mg/L为最优的遗传转化方案,转化率最高可达35.7%。另外,GUS瞬间表达率和转化率并不存在绝对的相关性,但瞬间表达分析仍然可以作为外源基因进入受体细胞的指示。花椰菜农杆菌介导转化方案的优化研究为芸薹属蔬菜高效遗传转化提供了技术保障,有利于芸薹属蔬菜遗传育种与种质创新研究。     

Purification and characterization of peroxidase from cauliflower (Brassica oleracea L. var. botrytis) buds

Peroxidases (EC 1.11.1.7; donor: hydrogen peroxide oxidoreductase) are part of a large group of enzymes. In this study, peroxidase, a primer antioxidant enzyme, was purified with 19.3 fold and 0.2% efficiency from cauliflower (Brassica oleracea L.) by ammonium sulphate precipitation, dialysis, CM-Sephadex ion-exchange chromatography and Sephadex G-25 purification steps. The substrate specificity of peroxidase was investigated using 2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulphonic acid) (ABTS), 2-methoxyphenol (guaiacol), 1,2-dihydroxybenzene (catechol), 1,2,3-trihyidroxybenzene (pyrogallol) and 4-methylcatechol. Also, optimum pH, optimum temperature, optimum ionic strength, stable pH, stable temperature, thermal inactivation conditions were determined for guaiacol/H(2)O(2), pyrogallol/H(2)O(2), ABTS/H(2)O(2), catechol/H(2)O(2) and 4-methyl catechol/H(2)O(2) substrate patterns. The molecular weight (M(w)) of this enzyme was found to be 44 kDa by gel filtration chromatography method. Native polyacrylamide gel electrophoresis (PAGE) was performed for isoenzyme determination and a single band was observed. K(m) and V(max) values were calculated from Lineweaver-Burk graph for each substrate patterns.     

Transformation of cauliflower (Brassica oleracea L. var. botrytis) — an experimental survey

The paper compares different approaches for the genetic transformation of cauliflower (Agrobacterium-mediated, PEG-mediated and/or electroporation). Transient expression of the neomycin phosphotransferase II (NPTII) gene could be detected after direct gene transfer. Stable transformation was achieved using both Agrobacterium-mediated and direct gene transfer. Expression as well as incorporation of the NPTII sequence could be demonstrated.     

The chromoplasts of Or mutants of cauliflower (Brassica oleracea L. var. botrytis)

Summary. The Or mutation in cauliflower (Brassica oleracea L. var. botrytis) leads to abnormal accumulations of -carotene in orange chromoplasts, in tissues in which leucoplasts are characteristic of wild-type plants. Or chromoplasts were investigated by light microscopy of fresh materials and electron microscopy of glutaraldehyde- and potassium permanganate-fixed materials. Carotenoid inclusions in Or chromoplasts resemble those found in carrot root chromoplasts in their optical activity and angular shape. Electron microscopy revealed that the inclusions are made up of parallel, membrane-bound compartments. These stacks of membranes are variously rolled and folded into three-dimensional objects. We classify Or chromoplasts as membranous chromoplasts. The Or mutation also limits plastid replication so that a single chromoplast constitutes the plastidome in most of the affected cells. There are one to two chromoplasts in each cell of a shoot apex. The ability of differentiated chromoplasts to divide in the apical meristems of Or mutant plants resembles the ability of proplastids to maintain plastid continuity from cell to cell in meristems of Arabidopsis thaliana mutants in which plastid replication is drastically limited. The findings are used to discuss the number of levels of regulation involved in plastid replication.     

Temporal and spatial root development of cauliflower (Brassica oleracea L. var. botrytis L.)

Row crops are often inefficient in utilizing soil resources. One reason for this appears to be inefficient rooting of the available soil volume. Five experiments were performed to study the temporal and spatial root development of cauliflower (cv. Plana). The crop was grown with 60 cm between rows, and root development was followed in minirhizotrons placed under the crop rows, 15 cm, and 30 cm from the crop rows. Soil was sampled and analyzed for nitrate content at the final harvest and once during growth. In two of the experiments N fertilizer rate was varied and in two of the other experiments two cultivars were compared (cv. Plana and Siria).The rooting depth of cauliflower was found to be linearly related to temperature sum, with a growth rate of 1.02 mm day^-1 °C^-1. Depending on duration of growth this leads to rooting depths at harvest of 85–115 cm. Soil analysis showed that the cauliflower was able to utilize soil nitrogen down to at least 100 cm.With Plana differences in root growth between row and interrow soil were only observed during early growth, but with Siria this difference was maintained until harvest. However, at harvest both cultivars had depleted row and interrow soil nitrate equally efficient. Nitrogen fertilizer did not affect overall root development significantly.The branching frequency of actively branching roots was increased in all soil layers from about 6 to 10 branches cm^-1 by increasing N fertilizer additions from 130 to 290 kg N ha^-1. Increasing N supply increased the number of actively branching roots in the topsoil and reduced it in the subsoil.The average growth rate of the roots was always highest in the newly rooted soil layers, but fell during time. At 74 days after planting very few roots were growing in the upper 60 cm of the soil whereas 70% of the root tips observed in the 80–100 cm soil layer were actively growing. Within each soil layer there was a large variation in growth rate of individual root tips.     

GA_3对老化花椰菜种子活力和几种相关生理生化性状的影响

用10、50、100、150和200mg·L-15种浓度赤霉素（GA3）溶液处理于10℃冰箱中贮藏5年的花椰菜老化种子。结果表明,100mg·L-1GA3浸种22h的效果最佳,老化种子的发芽率、发芽势、活力指数、根长均有提高,畸形苗率下降;过氧化物酶（POD）、过氧化氢酶（CAT）和脱氢酶活性提高,可溶性蛋白和叶绿素含量也提高,种子浸出液的电导率和丙二醛（MDA）含量则下降。另外,100mg·L-1GA3处理后的种子在温度为25℃条件下贮藏时间不宜超过25d。     

The Interaction of Plant Growth Regulators and Vernalization on the Growth and Flowering of Cauliflower (Brassica oleracea var. botrytis)

The growth and flowering response of a cold-requiring cauliflower (Brassica oleracea var. botrytis cv. 60 day) to a range of temperatures under 10 h photoperiod and to growth regulator application were investigated. Endogenous gibberellin A₁(GA₁) concentrations were also assessed under these treatments. Flowering and growth of the inflorescence stalk were correlated with plant developmental stage at the time of a vernalizing cold treatment. Temperature and its duration also affected flowering and inflorescence development. The most effective temperature for inflorescence induction was 10 °C. Flowering did not occur in non-vernalized plants (25 °C) even though they had been treated with GA₃. Application of GA₃ promoted inflorescence stalk elongation greatly in vernalized plants (10 °C), but less so in partially vernalized plants (15 °C or 20 °C). Paclobutrazol (PP₃₃₃) sprayed at the 8–9 leaf stage significantly suppressed inflorescence stalk length and slightly delayed flower bud formation and anthesis. Vernalization at 10 °C increased endogenous GA₁ content in both leaves and the inflorescence stalk irrespective of GA₃ or PP₃₃₃ treatment. Application of GA₃ tended to increase GA₁ levels, while PP₃₃₃ significantly reduce GA₁, both irrespective of vernalization. Vernalization is an important factor for flowering, but not curd formation in this cauliflower cv. 60 day and GA₁ is likely a causal factor in inflorescence stalk elongation.     

Cauliflower (Brassica oleracea L.) Seed Vigour: Imbibition Effects

By means of a machine vision facility, the process of water-imbibitionin a small seeded brassica species was recorded as the visibleincrease in seed volume. Dry cauliflower seed (Brassica oleracea)showed an immediate rapid phase of imbibition upon the additionof water. This initial phase was associated with the reductionin seedling root growth resulting from the imbibition of coldwater, and the rate of uptake of water at 20?C was negativelycorrelated with subsequent seedling growth. Damage to the testaof dry seed resulted in an increased rate of imbibition anda corresponding decrease in seedling root growth measured onslant boards. This showed that the intact testa of cauliflowerseed is capable of acting as a barrier to water influx and thata high rate of water uptake is damaging to the embryo. Testadamage reduced percentage soil emergence of seeds, as did raisingthe soil moisture content during the imbibition period by wateringimmediately after sowing. Both these treatments were believedto increase imbibition rate. Conditions which encouraged a lowrate of water uptake also improved the rate and uniformity ofemergence. Correlation of mean imbibition rates (measured ina laboratory test) with soil-emergence indicated that the sensitivityto imbibition damage varied between seed lots and interactedwith the absolute rate of water influx to determine the finalpercentage emergence. This factor prevented reliable predictionof seedling performance from the ranked-order of measured imbibitionrates. The significance of these findings to the seed productionand modular transplant raising industries is discussed. Key words: Machine vision, imbibition rate, testa, soil moisture, vigour     

Tissue-related changes in methyl-esterification of pectic polysaccharides in cauliflower (Brassica oleracea L. var. botrytis) stems

Pectic substances are a major component of cell walls in vegetable plants and have an important influence on plant food texture. Cauliflower (Brassica oleracea L. var. botrytis) stem sections at different regions of the mature plant stem have been monitored for tissue-related changes in the native pectic polysaccharides. Chemical analysis detected appreciable differences in the degree of methyl-esterification (ME) of pectic polysaccharides. About 65% of galacturonic acid (GalpA) residues were methyl-esterified in floret tissues. Relative ME showed a basipetal decrease, from 94% in the upper stem to 51% in the lower-stem vascular tissues. The decrease was not related to a basipetal increase in glucuronic acid (GlcpA) residues. The monoclonal antibodies, JIM 5 and JIM 7, produced distinct labelling patterns for the relatively low-methyl-esterified and high-methyl-esterified pectin epitopes, respectively. Labelling was related to cell type and tissue location in the stem. Floret cell walls contained epitopes for both JIM 5 and JIM 7 throughout the wall. Stem vascular tissues labelled more strongly with JIM 5. Whereas pith parenchyma in the upper stem labelled more strongly with JIM 7, in the lower-stem pith parenchyma, JIM 5 labelling predominated. Localization of pectic polysaccharide epitopes in cell walls provides an insight into how structural modifications might relate to the textural and nutritional properties of cell walls. Received: 16 August 1997 / Accepted: 20 December 1997     

Plant regeneration from cotyledonary protoplasts of cauliflower (Brassica oleracea L. var.botrytis L.)

Summary Protoplasts isolated enzymatically from precultured cotyledonary leaves ofB. oleracea var.botrytis and cultured in KM8p medium (Kao andMichayluk 1975) underwent sustained divisions in about 0.1% population to eventually produce callus, whereas mesophyll protoplasts from either field grown orin vitro raised plants failed to divide. The callus readily differentiated on Murashige-Skoog medium as modified for shoot culture (Binding 1974) to give rise to shoot and roots.     

Root growth of cauliflower (Brassica oleracea L. botrytis) under unstressed conditions: Measurement and modelling

Plant and Soil - Root observations were carried out on cauliflower using the minirhizotron and the soil core method in two years on two locations with different soil types, a loess loam and a humic...     

A structure-specific endonuclease from cauliflower (Brassica oleracea var. botrytis) inflorescence.

A protein with structure-specific endonuclease activity has been purified to near homogeneity from cauliflower ( Brassica oleracea var. botrytis) inflorescence through five successive column chromatographies. The protein is a single polypeptide with a molecular mass of 40 kDa. Using three different branched DNA structures (flap, pseudo-Y and stem-loop) we found that the enzyme, a cauliflower structure-specific endonuclease, cleaved the single-stranded tail in the 5'-flap and 5'-pseudo-Y structures, whereas it could not incise the 3'-flap and 3'-pseudo-Y structures. The incision points occur around the single strand-duplex junction in these DNA substrates and the enzyme leaves 5'-PO4 and 3'-OH termini on DNA. The protein also endonucleolytically cleaves on the 3'-side of the single-stranded region at the junction of unpaired and duplex DNA in the stem-loop structure. The structure-specific endonuclease activity is stimulated by Mg2+ and by Mn2+, but not by Ca2+. Like mammalian FEN-1, the protein has weak 5'-->3' double-stranded DNA-specific exonuclease activity. These results indicate that the cauliflower protein is a plant structure-specific endonuclease like mammalian FEN-1 or may be the plant alternative.