Plant regeneration from mesophyll protoplasts of Moricandia arvensis

Moricandia arvensis is of interest as it is a dicotyledonous species which has C₃ — C₄ intermediate photosynthesis, a mechanism which results in enhanced recapture of photorespired CO₂. Leaves from cultured shoot tips were used as a source for mesophyll cell protoplasts. Approximately 1% of the protoplasts which survived the first few days of culture produced calli. On a suitable regeneration medium, 30–60% of the calli regenerated one or more shoots. From among the regenerating shoots eight were selected, transferred to soil and grown to flowering in the glasshouse; all were fertile. The development of a protoplast regeneration system provides the opportunity to use transformation and somaclonal variation as tools in the genetic analysis of the C₃–C₄ character in this species.Abbreviations GDC glycine decarboxylase - 2,4-D 2,4-dichlorophenoxyacetic acid - BAP benzyl aminopurine - NAA naphthalene acetic acid - ABA abscisic acid     

紫花苜蓿叶输导组织解剖结构和光合特性对外源氮的响应

以紫花苜蓿品种‘甘农3号’和‘陇东苜蓿’为研究材料,采用室外(防雨网室)盆栽营养液砂培法,研究了2种氮素形态(NO3--N,NH4+-N)的5个氮素水平(0、105、210、315、420mg·L-1)处理对叶片输导组织解剖结构及光合特性的影响。结果显示:(1)与无氮处理相比,供氮处理下‘甘农3号’和‘陇东苜蓿’输导组织的解剖结构和光合特性发生明显变化,叶片输导组织维管束、木质部和韧皮部面积显著增大,导管数显著增多,净光合速率、气孔导度、叶面积和叶绿素含量均显著增加,且在210mg·L-1供氮水平下达到最大值。(2)2种氮素形态相比,‘甘农3号’和‘陇东苜蓿’叶片输导组织维管束面积、木质部面积、韧皮部面积、导管数、净光合速率、气孔导度、叶面积、叶绿素含量均表现为NH4+-N处理好于NO3--N处理。(3)2品种相比,叶片输导组织维管束面积、木质部面积、韧皮部面积、导管数、净光合速率、气孔导度、叶面积、叶绿素含量表现为‘甘农3号’大于‘陇东苜蓿’。研究表明,氮素能通过改善紫花苜蓿叶片输导组织的解剖结构和光合特性,促进紫花苜蓿光合作用;各处理中以NH4+-N、210mg·L-1表现最佳,维管束面积最大,木质部、韧皮部面积大且发育好,导管数最多,而且‘甘农3号’表现更优。     

不同倍性大青杨的光合特性及叶片解剖结构比较

通过对二年生二倍体、三倍体和四倍体大青杨叶片结构和光合特性的研究,探讨不同倍性大青杨生长性状差异的原因。结果表明：不同倍性大青杨的净光合速率（Pn）、光饱和点（LSP）、光补偿点（LCP）等光合指标差异显著。三倍体的最大净光合速率（Pmax）高出二倍体21%。叶片厚度、表皮细胞厚度、栅栏组织厚度、栅栏组织与海绵组织厚度的比值,都是三倍体和四倍体大青杨比二倍体高,且均呈现增加的趋势。其中,四倍体大青杨表皮细胞最厚,上下表皮分别高出二倍体64%和17%。三倍体大青杨的栅栏组织厚度及栅栏组织与海绵组织厚度的比值最大,比值高出二倍体50%。通过叶片光合特性和解剖结构比较证明,三倍体和四倍体大青杨对光的适应性和光合作用能力强于二倍体。     

Somatic embryogenesis and plant regeneration from stem explants of Moricandia arvensis

Efficient and reproducible plant regeneration has been established from stem internode explants of Moricandia arvensis, a crucifer of special interest due to its C₃-C₄ intermediate photosynthetic activity. Somatic embryogenesis was induced in one-third of explants cultured on Murashige and Skoog based medium containing 9 mm 2,4-dichlorophenoxyacetic acid. High frequencies of plant regeneration (>90%) resulted when somatic embryos were germinated on medium lacking growth regulators. Regenerated plants were diploid, fertile and morphologically similar to seed-derived plants of M. arvensis. This is the first report of somatic embryogenesis in M. arvensis. This plant regeneration system should facilitate gene identification and localisation studies of C₃-C₄ physiology by insertional mutagenesis, a prerequisite for the isolation and transfer of genes involved in C₃-C₄ metabolism from Moricandia to cultivated brassicas. Received: 28 November 1996 / Revision received: 30 January 1997 / Accepted: 15 February 1997     

Somatic hybridization between Brassica juncea and Moricandia arvensis by protoplast fusion

Intergeneric somatic hybrids have been produced between Brassica juncea (2n=36, AABB) cv. RLM-198 and Moricandia arvensis (2n=28, MM) by protoplast fusion. Hypocotyl protoplasts of B. juncea were fused with mesophyll protoplasts of M. arvensis using polyethylene glycol. Fusion frequency, estimated on the basis of differential morphological characterstics of parental protoplasts was about 5%. Of the 156 calli obtained, four calli produced shoots intermediate in morphology between the parents. Hybrid nature of the plants was confirmed using wheat nuclear rDNA probe. Hybridization of total DNA with a mitochondrial DNA probe carrying 5s–18s rRNA genes of maize showed that the mitochondria of the somatic hybrids were derived from the wild species M. arvensis. Meiosis in the only hybrid that produced normal flowers revealed the occurrence of 64 chromosomes, the sum of chromosomes of parental species. Inspite of complete pollen sterility, siliquas were produced in this hybrid by back-crossing with B. juncea. These siliquas on in vitro culture produced 12 seeds.     

Influence of Genome on Leaf Anatomy of Triticum and Aegilops

The leaf mesophyll of Triticum and Aegilops is constructed fromcells with one to ten arms. Volume of mesophyll cells per unitleaf area was larger in some monogenomic (A and B genome) plantsthan in polyploids, while leaf volume per unit leaf area wassmaller in the former than in the latter. Consequently, thecompactness of leaf blade is higher in these monogenomic plantsthan in the polyploids. D genome plants showed a much lowervolume of both mesophyll cells and leaf blade per unit leafarea, but the compactness of the leaf blade was generally higherthan in the polyploids. Mesophyll surface area per unit leaf area tended to be largerin the A and B genome than in the D genome and polyploid plants.Out of the polyploids, AB genome plants showed a larger mesophyllsurface area per unit leaf area as compared with AG and ABDgenome plants. Therefore, either the D or the G genome seemsto have the effect of decreasing the mesophyll surface areaper unit leaf area. A decrease of the compactness of leaf bladeand the mesophyll surface area per unit leaf area in the polyploidswas considered to be associated with the reduction of theirphysiological activities on the unit leaf area basis. Triticum, Aegilops, wheat, mesophyll surface area, leaf anatomy, genome, photosynthesis     

Photorespiratory ammonia assimilation in the leaves of barley, sorghum and Moricandia arvensis

Effects of methionine sulphoximine (MSX) on glutamine synthetase activity and ammonia accumulation in the leaves of barley, sorghum and Moricandia arvensis were studied. Glutamine synthetase activity of sorghum leaves was relatively resistant to MSX treatment during initial stages. All the three plant species exhibited considerable accumulation of ammonia arising from non-photorespiratory sources, in response to MSX treatment. Minimal estimates of the rate of glycine decarboxylation were made employing MSX and isonicotinyl hydrazide together in the treatment solution. Changes in the pool sizes of amino acids involved in photorespiratory nitrogen metabolism as affected by MSX have also been investigated.     

Transgenic plant production from leaf discs of Moricandia arvensis using Agrobacterium tumefaciens

Summary A high frequency shoot regeneration (80%) was developed from callus of leaf discs and stem internodes of Moricandia arvensis. Leaf discs were shown to be a preferable starting material for transformation experiments. Agrobacterium tumefaciens strain GV3101/pMP90 used in this study contained a binary vector with genes for kanamycin resistance, hygromycin resistance and -glucuronidase (GUS). Maximum transformation efficiency (10.3%) was achieved by using kanamycin at the rate of 200 mg/l as a selection agent. Presence of tobacco suspension culture during co-cultivation and a pre-selection period of seven days after co-cultivation was essential for successful transformation. Transgenic plants grew to maturity and exhibited flowering in a glasshouse. GUS activity was evident in all parts of leaf and the presence of GUS gene in plant gemone was confirmed by PCR analysis.Abbreviations GUS -glucuronidase     

Photosynthetic/Photorespiratory Carbon Metabolism in the C(3)-C(4) Intermediate Species, Moricandia arvensis and Panicum milioides

The distribution of ¹⁴C in photosynthetic metabolites of two naturally occurring higher plants with reduced photorespiration, Moricandia arvensis and Panicum milioides, in pulse and pulse-chase ¹⁴CO₂ incorporation experiments was similar to that for the C₃ species, M. foetida and Glycine max. After 6 seconds of ¹⁴CO₂ incorporation, only about 6% of the total ¹⁴C fixed was in malate and aspartate in both M. arvensis and P. milioides. The apparent turnover of the C₄ acids was very slow, and malate accumulated during the day in M. arvensis. Thus, C₄ acid metabolism by M. arvensis and P. milioides had no significant role in photosynthetic carbon assimilation under the conditions of our experiments (310 microliters CO₂ per liter, 21% O₂, 1100 or 1900 micromoles photon per square meter per second, 27°C).

After a 36-second chase period in air containing 270 microliters CO₂ per liter, about 20% of the total ¹⁴C fixed was in glycine with M. arvensis, as compared to 15% with M. foetida, 14% with P. milioides, and 9% with G. max. After a 36-second chase period in 100 microliters CO₂ per liter, the percentage in glycine was about twice that at 270 microliters CO₂ per liter in the C₃ species and P. milioides, but only 20% more ¹⁴C was in glycine in M. arvensis. These data suggest that either the photorespiratory glycine pool in M. arvensis is larger than in the other species examined or the apparent turnover rate of glycine and the flow of carbon into glycine during photorespiration are less in M. arvensis. An unusual glycine metabolism in M. arvensis may be linked to the mechanism of photorespiratory reduction in this crucifer.

     

Flag Leaf Anatomy of Triticum and Aegilops Species in Relation to Photosynthetic Rate

Quantitative anatomical and other measurements were made onfully expanded flag leaves of a series of diploid, tetraploidand hexaploid Triticum and Aegilops species, and photosyntheticrates per unit leaf area were measured at light saturation (P_max). Diploids had the highest P_max, hexaploids the lowest with tetraploidsbeing intermediate. The anatomical features of tetraploids andhexaploids were generally similar, but different from the diploids.The diploids had thinner leaves with less dry matter and chlorophyllper unit area. The surface area of the mesophyll cells per unitvolume of mesophyll tissue was similar for all ploidy levels,as was the ratio mesophyil cell surface area per unit leaf area.It is argued that while these anatomical features are unlikelyto account for the observed variation in Pmax, it is possiblethat other structural factors with which they are correlatedmay causally influence P_max. One such feature is the averagediffusion path length from the plasmalemma at the cell surfaceto the sites of carboxylation. Anatomy, photosynthesis, mesophyll, cell size, Triticum, Aegilops, polyploidy     

5种叶面肥对草莓光合特性及品质的影响

为探讨不同叶面肥对大棚草莓光合特性、果实品质及产量的影响,并筛选出适宜的叶面肥,以甘肃省兰州市西固区牟家台草莓温室大棚内栽植的草莓品种‘蒙特瑞’为试材,在施肥充足的情况下,在4个生育期追肥喷施经典750倍液（T1）、花仆1500倍液（T2)、润亮750倍液(T3)、艾德拉果500倍液（T4）及磷酸二氢钾1500倍液（T5）5种叶面肥,清水作为对照（CK）,测定了处理植株光合参数、果实品质、生物量及产量等指标。结果表明：（1）喷施不同叶面肥处理后草莓植株生物量、叶绿素含量显著增加,叶片的净光合速率和蒸腾速率显著上升,喷施叶面肥后的草莓植株胞间二氧化碳浓度均高于CK;（2）对果实品质而言,T2和T3处理能够显著改善草莓果形指数,T1和T5可以显著增大果实体积和重量。其中,T1的平均单果重较CK增加了5.89 g,T5的果实横径和纵径分别较CK增加了1.22 cm和1.15 cm。（3）实硬度、可溶性固形物含量、维生素C含量和还原糖含量有很明显的提高,并以T1施用效果最佳,较CK分别增加了3.3%、17.25 mg/g、2.71%。处理T1和T3可溶性蛋白含量分别较CK提高了0.097 mg/g和0.070 mg/g;（4）喷施叶面肥能够显著提高草莓单位面积产量,且以处理T1处理产量最高,较CK增长了13.65%。可见,施用适宜的叶面肥均能有效改善草莓营养生长状况,提升草莓果实品质,显著提高产量,并以经典750倍液叶面肥施用综合效果最佳。     

8种绿化树种光合特性及叶片解剖结构比较

城市绿化不仅包含了园林绿化的美化作用,还具有重要的生态功能,其生态功能是通过植物的生理活动实现的。光合能力在种间和基因型间的变化很大,这些差异通常与代谢和(或)叶片的解剖结构的性质有关。本研究选择8种哈尔滨常见树种,采用Li-6400便携式光合测定系统对叶净光合速率(P_n)、呼吸速率(R_d)、蒸腾速率(T_r)、气孔导度(G_s)、胞间CO_2浓度(C_i)等进行测定,并利用显微镜观察测定叶片厚度、表皮厚度、栅栏组织厚度、海绵组织厚度,从而探讨叶片结构对光合生理的影响。结果表明:8个树种间叶片最大光合速率、气孔导度、胞间CO_2浓度、蒸腾速率、光饱和点差异显著(P <0.05);表皮厚度、栅栏组织厚度、上表皮气孔密度和下表皮气孔密度差异显著(P <0.05)。虽然8个树种间光合能力和叶片解剖结构的差异较大,但分析发现其间也存在一定的相关性。其中,光饱和点与叶表皮厚度显著正相关(P <0.01),相关系数为0.78。胞间CO_2浓度与上表皮气孔密度显著负相关(P <0.05),相关系数为-0.65。而最大光合速率、呼吸速率、蒸腾速率和光补偿点与表皮厚度、栅栏组织厚度、海绵组织厚度、上表皮气孔密度和下表皮气孔密度相关均不显著(P> 0.05)。胞间CO_2浓度与表皮厚度、栅栏组织厚度、海绵组织厚度和下表皮气孔密度相关均不显著(P> 0.05)。光饱和点与栅栏组织厚度、海绵组织厚度、上表皮气孔密度和下表皮气孔密度相关均不显著(P> 0.05)。虽然对叶片结构对生理过程的影响的机理还需要进一步研究,但是我们认为叶片解剖结构的研究可以更好地理解生理指标的变化。     

3个玉米黄叶突变体的光合特性研究

在农业生产中光合作用是作物积累生物量的主要方式,其主要依赖于多种光合色素和完整的叶绿体结构与功能。而玉米叶色突变体对于研究叶绿体发育、提高玉米光合作用能力和产量具有重要意义。以两个玉米自交系郑58(Z58)和B73为对照,对从甲基磺酸乙酯(ethyl methanesulphonate,EMS)处理后的不同玉米诱变群体中筛选到的2株黄叶突变体yl-1(yellow leaf-1,Z58背景)、yl-2(yellow leaf-2,B73背景)以及从玉米自交系Z58中发现的1株自然黄叶突变体yl-3(yellow leaf-3)等3个表型相似的玉米黄叶突变体的形态特征、光合色素含量、叶绿素合成前体物质含量进行了比较研究。结果表明,与对照相比,3个突变体在整个生长周期内均呈现不同程度的黄叶表型、不复绿、植株矮小、发育迟缓;叶片总叶绿素、叶绿素a和叶绿素b含量均显著降低(P<0.05),叶绿素a/叶绿素b比值显著升高(P<0.05);不同突变体的各类叶绿素合成前体物质含量有不同程度的降低。3个突变体的黄叶表型可能是由不同基因的突变导致相关四吡咯化合物合成异常引起的。研究结果为定位...     

Intergeneric (intersubtribe) hybridization between Moricandia arvensis and Brassica A and B genome species by ovary culture

Summary Intergeneric hybrids between Moricandia arvensis (C₃–C₄ intermediate species) and Brassica A and B genome species (B. campestris and B. nigra) were produced via ovary culture. When M. arvensis was used as a female parent, the hybrid embryo yield (0.25–0.45 embryo per pollination) was similar between two genomes, regardless of the male parent. The reciprocal hybrid using B. campestris as a female was also obtained, although yield of embryo was lower (0.02 embryo per pollination). On the other hand, no hybrids were obtained without the in vitro technique. As most hybrid embryos could not develop normal shoots, plants were regenerated by inducing shoots on the cultured hypocotyl. The hybrid nature of the regenerated plant was confirmed morphologically and cytogenetically. A certain amount of bivalents (2.52-2.71) in the hybrids indicated the existence of partial chromosome homology between two genera. The present results indicate that ovary culture is an effective technique for overcoming the crossing barrier between M. arvensis and Brassica cultivated species.     

遮阴对猫爪草光合特性及叶片解剖结构的影响

【目的】探究不同遮阴度对野生植物猫爪草（Ranunculus ternatus Thunb）光合特性和叶片解剖结构的影响,为深入了解其耐阴性以及开展规模化栽植提供理论基础。【方法】采用控制变量法,以盆栽猫爪草为试验材料,设置全光照（CK）、30%遮阴（T1）、50%遮阴（T2）、70%遮阴（T3）和90%遮阴（T4）5个处理,对不同遮阴处理下的光合特性及叶片解剖结构进行分析。【结果】（1）随着遮阴程度增加,叶面积在T3环境下达到最大值,株高和茎粗逐渐降低。（2）随遮阴程度的增加,叶片表观量子效率（AQE）先升后降,在T4处理下显著低于其他处理,而光饱和点（LSP）、光补偿点（LCP）、暗呼吸速率（Rd）和最大净光合速率（Pnmax）逐渐降低;最大荧光（Fm）、可变荧光（Fv）、最大化学效率（Fv/Fm）和PSⅡ潜在活性（Fv/F0）先升后降,初始荧光（F0）热耗散的电子比率（φDo）则先降后升。（3）随着遮阴程度的增加,叶片气孔密度先减后增,并在T3处理达到最小值,上下表皮厚度和海绵组织厚度逐渐降低,组织结构紧密度（CTR）先降后升,组织结构疏松度（SR）先升后降。（4）叶片栅栏组织、海绵组织、叶片厚度及上、下表皮厚度与Rd呈极显著或显著正相关关系;海绵组织厚度、CTR和SR与Pnmax ,以及栅海比（P/S）、SR与AQE均呈显著正相关关系;海绵组织厚度和SR与F0呈显著负相关关系。【结论】猫爪草可以通过降低光合速率和改变叶片结构来提高耐阴性,在全光照下生长良好,也能较好地适应30%-70%遮阴环境。     

Effects of Long-Term Fertilization on Leaf Photosynthetic Characteristics and Grain Yield in Winter Wheat

Based on a 20-year fertilization experiment with wheat-maize double cropping system, the effects of different long-term fertilization treatments on leaf photosynthetic characteristics and grain yield in different winter wheat (Triticum aestivum L.) cultivars were studied in the growing seasons of 2000–2001 and 2001–2002. A total of nine fertilization treatments were implemented, i.e. no fertilizer (CK), N fertilizer (N), N and P fertilizers (NP), N and K fertilizers (NK), N, P, and K fertilizers (NPK), only organic manure (M), organic manure and N fertilizer (MN), organic manure and N and P fertilizers (MNP), and organic manure and N, P, and K fertilizers (MNPK). With the treatments of combined organic manure and inorganic fertilizers (TMI), net photosynthetic rate (P _N), maximal activity of photosystem 2, PS2 (F_v/F_m), and chlorophyll content (SPAD value) of flag leaves and leaf area index (LAI) were much higher at the mid grain filling stage (20 or 23 d post anthesis, DPA), and exhibited slower declines at the late grain filling stage (30 DPA), compared with the treatments of only inorganic fertilizers (TI). The maximal canopy photosynthetic traits expressed as P _N×LAI and SPAD×LAI at the mid grain filling stage were also higher in TMI than those in TI, which resulted in different grain yields in TMI and TI. Among the treatments of TMI or among the treatments of TI, both flag leaf and canopy photosynthetic abilities and yield levels increased with the supplement of inorganic nutrients (N, P, and K fertilizers), except for the treatment of NK. Under NK, soil contents of N and K increased while that of P decreased. Hence the unbalanced nutrients in soil from the improper input of nutrients in NK treatment were probably responsible for the reduced flag leaf and canopy photosynthetic characteristics and LAI, and for the fast declining of flag leaf photosynthetic traits during grain filling, resulting in the reduced yield of NK similar to the level of CK.     

硝态氮对海水胁迫下长春花幼苗光合特性及离子含量的影响

采用砂培试验研究了硝态氮对海水处理下长春花幼苗光合特性和离子分布的影响.结果表明：（1）高浓度海水显著抑制长春花幼苗的生长,提高硝态氮浓度能显著缓解海水胁迫对长春花幼苗生长发育的抑制作用,促进地上、地下部生物量积累.（2）随着海水浓度的增加,长春花幼苗叶片净光合速率、蒸腾速率、气孔导度和水分利用效率较对照均显著降低,细胞间隙CO2浓度显著增加;增加硝态氮浓度可显著提高长春花幼苗的净光合速率、蒸腾速率、气孔导度和水分利用效率,显著降低细胞间隙CO2浓度.（3）随着海水浓度的增加,长春花幼苗体内K＋含量较对照显著降低,而Na^＋、Cl^-含量显著增加,提高硝态氮浓度有利于长春花幼苗对K＋的吸收和向上运输,维持地上部较高的K^＋/Na^＋.研究发现,增加硝态氮能维持高浓度海水胁迫下长春花幼苗体内养分平衡,缓解盐胁迫对其光合生理和生长造成的伤害.     

Effects of Elevated Atmospheric CO2 Concentration on Leaf Anatomy and Morphology in Panicum Species Representing Different Photosynthetic Modes

Panicum tricanthum Nees, Panicum antidotale Retz., and Panicum decipiens Nees ex Trin. were selected to represent C3, C4, and C3/C4 intermediate perennial species of Panicum, respectively. Plants grown from seed with 900 ppm [CO2] under natural sunlight and controlled temperatures (30 degrees /22 degrees C) were compared with plants grown with ambient [CO2]. The anatomy of the last fully expanded leaf of the main tiller was studied by light microscopy with computerized graphic image analysis and by transmission electron microscopy. Leaf anatomy did not change qualitatively in response to elevated [CO2], but there were changes in leaf thickness and in the proportions of total transsectional area occupied by mesophyll, bundle sheath cells, vascular elements, and sclerenchyma, according to species. The abaxial stomatal frequency decreased by 22% for P. tricanthum but increased by ca. 30% for the other two species. With 900 ppm CO2, all three species showed a considerable increase in leaf starch content (to >30% of dry matter). Starch granules accumulated in chloroplasts of the mesophyll and bundle sheath cells. Increased leaf glaucousness in response to elevated [CO2] was the result of increased or modified deposition of epicuticular wax on both leaf surfaces, a response to elevated [CO2] that is unusual and one that has not been previously recorded for monocotyledons. The wax patterns were studied by scanning electron microscopy. Panicum decipiens did not respond to elevated [CO2] in a truly intermediate fashion; its responses resembled those of either the C3 or the C4 species. C3/C4 intermediates may thus be interpreted as developmental chimeras and not as species in transition between C3 and C4 modes in an evolutionary sense.