Differences in response to the toxin sirodesmin PL produced by Phoma lingam (Tode ex fr.) Desm. on protoplasts,cell aggregates and intact plants of resistant and susceptible Brassica accessions

Summary The selective property of sirodesmin PL, a toxin produced by Phoma lingam, was studied on protoplasts, cell aggregates, leaves and roots. Directly after isolation, protoplasts from all the different Brassica accessions were sensitive when treated with toxin in a concentration higher than 1 M. When more differentiated plant tissue. i.e. cell aggregates, leaves or roots, were investigated, insensitivity to the toxin was found in the plant material resistant to P. lingam, while the plant material susceptible to P. lingam was sensitive. The results reveal that a clear correlation between resistance to P. lingam and insensitivity to sirodesmin PL is present, and that the toxin can be used to distinguish resistant plant material from susceptible both in vivo and in vitro.     

High Yield Isolation of Nuclei from Plant Protoplasts

Nuclei were isolated from protoplasts obtained from Parthenocissus tricuspidata Crown Gall callus tissues. The effect of various isolation procedures, detergent or ultrasonication, on yield and quality of nuclei was studied. A standard procedure, based on the use of 5 × 10^?3% Triton × 100 — 6% PVP — 20% glycerol, may be carried out in 30 min and gives 80 to 90% yield of nuclei in which RNA polymerase activity is retained.     

Isolation of active DNA-binding nuclear proteins from tomato galls induced by root-knot nematodes

We describe 2 methods for extraction of DNA-binding proteins from root-knot nematode feeding sites (ie, galls). DNA-binding activity was assayed by electrophoretic mobility shift assays using fragments from the root-knot nematode-responsiveLEMMI9 and 35S promoters. In noninfected tissue, the method based on nuclei enrichment through a Percoll cushion was superior for isolation of DNA-protein binding activity with both promoters. With infected roots; the method based on crude extracts performed better with theLEMMI9 promoter, whereas nuclei-enriched extracts worked better with the 35S promoter. Therefore, both methods can be used to extract proteins for DNA-binding assays from infected roots, but the method of choice may depend on the promoter under study. Both authors have contributed equally to this work.     

Effects of feeding by larvae of Phaedon cochleariae (F.) and Plutella maculipennis (Curt.) on the yield of radish and turnip plants

Different numbers of larvae of the beetle Phaedon cochleariae (F.) and the moth Plutella maculipennis (Curt.) were fed on the leaves of turnip plants (variety Early Milan White) and radish (variety French Breakfast) growing in a constant-environment room. Turnips grew more slowly than radish, were less affected by insect feeding and were affected more by Phaedon than by Plutella. Phaedon larvae severed leaf veins and rasped the leaf surface, thus killing more leaf tissue by desiccation than they ate. They remained on and greatly damaged the older leaves and their feeding reduced yield. Plutella larvae fed on leaves of all ages, which they ate cleanly, leaving the veins. They pupated sooner than Phaedon and did less damage to older leaves, which grew larger and lived longer than corresponding leaves on intact plants. This compensatory growth by turnip prevented loss of yield and may have increased it, as indicated by dry weight of the roots. Both species of insect decreased the yield of radish similarly. Loss of yield of turnip caused by Phaedon or of radish caused by both species of insects was negatively and linearly proportional to the number of larvae that fed on the leaves.     

Nuclear DNA content of Pinus sylvestris (L.) as determined by laser flow cytometry

Nuclear DNA content was determined in nuclei isolated from needles, stems and roots of in vitro grown seedlings and from megagametophytes and embryo of mature seeds in three accessions of Pinus sylvestris L. One accession was from Inari, northern Finland at timber line, and two accessions were from the Alpine region in Italy. Nuclei were mechanically isolated by a chopping method, stained with propidium iodide, and DNA content was determined using an EPICS PROFILE laser flow cytometer. Nuclei isolated from leaves of barley (Hordeum vulgare L. cv. Sultan; 2C=11.12 pg) were used as an internal standard for measurement of pine nuclei. Mean 1C nuclear DNA content of P. sylvestris was 27.88 pg as determined from megagametophyte tissue. Mean 2C value was 52.25 pg as determined from stem and root tissue, and 55.58 pg as determined from embryo tissue. The ratio of 2C to 1C value was 1.87 and 1.99, respectively. Extracts of nuclei from needles contained propidium iodide-absorbing debris which may have interfered with measurements and resulted in lower 2C values than those obtained from stem and root.     

不同施磷水平对冷浸田水稻磷含量、光合特性及产量的影响

为探讨施磷对冷浸田水稻(Oryza sativa)生长的作用,研究不同施磷水平对水稻植株的磷含量、光合特性及产量的影响。结果表明,与对照相比,分蘖期增施磷肥使根、茎叶的磷含量和叶片净光合速率增加10.53%~36.84%、18.52%~37.03%和15.17%~29.88%;抽穗期使根、茎叶和稻穗的磷含量增加11.76%~23.53%、24.13%~41.38%和9.68%~22.58%,叶片净光合速率和可溶性糖含量提高13.41%~27.10%和6.03%~22.38%,而淀粉含量则降低12.05%~41.77%;成熟期的稻穗磷含量和产量增加0.71%~28.57%和4.65%~10.32%。施磷量(X)与稻谷产量(Y)满足方程:Y=-0.0835X~2+14.224X+6530.9(r=0.94*),抽穗期茎叶磷含量、叶片净光合速率与稻谷产量的相关系数分别为0.57*和0.77*。因此,冷浸田增施磷肥可提高水稻植株的磷含量、叶片净光合速率和可溶性糖含量,并降低叶片淀粉累积,从而提高稻谷产量。抽穗期是磷肥影响冷浸田水稻产量的关键时期,0.38%的茎叶磷含量可作为磷素养分丰缺的诊断指标,冷浸田水稻磷肥(P2O5)的推荐用量为85.17 kg hm–2。     

Light and ethylenediamine tetraacetic acid mediated differential effects of ammonium on nitrate reductase activity in maize seedlings

Abstract Effect of ammonium on in vivo activity of nitrate reductase in roots, shoots and leaves of maize (Zea mays L.) seedlings was studied in relation to light/dark conditions and EDTA supply. Supply of 5 mM (NH₄)₂SO₄ increased the steady state level of enzyme only in leaves and in light, while it had no effect in roots and shoots and in the dark. The substrate induction of enzyme was also little affected by 1 to 10 mM (NH₄)₂SO₄ in roots and shoots. In the leaves the activity in the dark was either inhibited (minus EDTA) or stimulated (plus EDTA) by 5 to 10 mM (NH₄)₂SO₄. The activity was stimulated in the light also in the presence of EDTA at higher concentrations of ammonium. When different concentrations of ammonium were supplied without any exogenous nitrate in the light, the enzyme activity increased at low concentration and was either inhibited or unaffected at higher concentrations depending upon the tissue used. Supply of EDTA with ammonium modified its effect to some extent. It is suggested that the effect of ammonium on nitrate reductase activity depends upon the tissue used and the effective concentration of the ammonium.     

The influence of ammonia in nutrient solution on growth and metabolism of cucumber plants

Summary Shoot yield of cucumber plants grown 18 days in nutrient solution with 0.06 mM NH₃ was decreased. Root yield was diminished at 0.09 mM NH₃ The ammonia treatment caused heavy chlorosis increasing with age of leaves. This chlorosis was not due to any nutrient deficiency. Ammonia also influenced the morphology of roots. They were clearly shorter caused by a much smaller size of root cells.The decrease of yield was linked to a reduction of assimilation occurring not only after a long influence of ammonia lasting 14 days, but also within one hour after starting the NH₃ treatment. The decline of assimilation was probably caused by a higher resistance of stomata against CO₂ influx in leaf tissue as can be concluded from the observation that transpiration was decreased in the same way as assimilation.The effect of ammonia in nutrient solution could also be due to the occurrence of higher NH₃ concentrations in leaf tissue, because both, pH of plant press sap as well as NH₄ concentration of plant tissue, were increased.Furthermore, it is shown that the nitrate content of plant tissue was diminished by ammonia whereas ammonium and amide content were raised. Regulation of nitrate uptake of plants by means of ammonium and amide content of tissue is discussed.     

Antitermitic effect of the Lantana camara plant on subterranean termites (Isoptera: Rhinotermitidae)

Abstract The insecticidal effects of Lantana camara L. (flowers, leaves, stems and roots) and the soil where lantana had been growing, on foraging activity and survival of the subterranean termites Coptotermes formosanus and Reticulitermes flavipes were examined in a 3-week experiment. The soil in which lantana had been growing had no effect on termite tunneling and survival. Incorporation of chipped fresh lantana leaves and stems into soil had no effect on mortality but caused significant reduction in tunneling. The 5-cm wide barrier of soil with lantana tissue incorporated effectively repelled groups of both species from penetrating the barrier and thus prevented infestation of a piece of wood on the other side of the barrier. C. formosanus was more sensitive in avoiding the barrier than R. flavipes. Leaves, stems and flowers were more repellent than roots. These results provide preliminary evidence that fresh-cut lantana leaves, stems and flowers may have use as additives to garden mulches against termites.     

Isolation of nuclei from rice tissue grown in suspension culture

Summary A simple method is described for the isolation of intact nucleic from rice tissue (Oryza sativa L.) grown in suspension culture. The procedure involves incubation of the tissue for 4 h with cellulase and pectinase prior to disruption of the cells. The yield of nucleic is approximately 40% (DNA basis) and the preparations are capable of synthesizing RNA in vitro. The method may be valuable to biochemically oriented research requiring plant nuclei.     

Effects of osmotic- and high-light stresses on PSII efficiency of attached and detached leaves of three tree species adapted to different water regimes

Abscisic acid (ABA), an important chemical signal from roots, causes physiological changes in leaves, including stomata closure and photoprotection. Furthermore, endogenous ABA concentration in leaves and stomatal behavior vary with the species adapted to different water regimes. In this study, Ficus microcarpa, a hemiepiphyte, Salix warburgii, a hygrophyte, and Acacia confusa, a mesophyte, were used to elucidate the effects of leaf detachment on photosystem II (PSII) efficiency under osmotic- and high-light stresses. Results indicate that, under osmotic- and high-light stresses, PSII efficiency of the detached leaves was lower than that of the attached leaves for all three tree species, when compared at the same levels of stomatal resistance and leaf water potential. Exogenous ABA could mitigate the PSII efficiency decrease of detached F. microcarpa leaves under osmotic- and high-light stresses. Yet, the osmotic stress could raise endogenous ABA concentration in the attached, but not in the detached F. microcarpa leaves. In addition, partial root-zone drying exerted a significant effect on stomatal behavior but not on the water status of F. microcarpa leaves. These observations imply that the stronger ability of PSII in the attached leaves of F. microcarpa under osmoticand high-light stresses was probably due to the protective action of ABA from roots. On the contrary, endogenous ABA level of S. warburgii leaves was very low. In addition, partial root-zone drying produced no significant effect on its stomatal behavior. Therefore, PSII in attached S. warburgii leaves was possibly protected from the damaging effects of excess absorbed energy by signals other than ABA, which were transported from the roots.     

Systemic Migration and Establishment of Xanthomonas campestris pv. pruni in Plum Leaves and Twigs

The systemic migration of Xanthomonas campestris pv. pruni (Xcp) through vascular bundles of leaves and twigs of plum was investigated. A rifampicin-resistant strain of Xcp was inoculated into leaves located midway from the tip of new green twigs of Golden King plum trees in a glasshouse. High numbers of the pathogen were recovered 4 and 8 weeks after inoculation from sections of uninoculated and symptomless veins, petioles, and twig tissue. Symptoms of bacterial spot developedwithin 8 weeks on main and secondary veins of uninoculated leaves located as far as 13 cm from the inoculated leaf on the same twig. Weekly isolation indicated the constant presence of Xcp in apparently unaffected shields of twig tissue obtained from a naturally infected Golden King orchard. Xcp apparently enters plum twigs through veins of infected leaves and migrates systemically through twigs to leaves.     

Effects of carrot psyllid (Trioza apicalis) feeding on carrot yield and content of sugars and phenolic compounds

Carrot psyllid, Trioza apicalis, is a serious pest of carrot in Northern Europe, as it can significantly damage young carrot seedlings in a period as short as 3 days. This study was conducted to investigate effects of carrot psyllid feeding at different plant growth stages on carrot yield and to assess changes in content of sugars, phenolics and related compounds in carrot roots resulting from the psyllid feeding. In addition, reflectance of carrot leaves was measured to assess the intensity of discolouration in damaged leaves. Results showed that carrot yield was significantly reduced by a 3‐day carrot psyllid feeding period when the seedlings were exposed to psyllids at 1‐ or 2‐leaf stage. However, at 4‐leaf stage feeding by one carrot psyllid did not reduce yield. Sucrose concentration in the damaged roots was significantly decreased, whereas concentrations of some phenolic compounds were significantly increased. The reflectance of leaves of damaged carrots differed significantly from those of undamaged control leaves. These observations indicate that carrot psyllid damage has potential to lower not only the carrot yield, but also the carrot crop quality. No phytoplasma was detected in the carrots exposed to psyllids, but recently, T. apicalis has been associated with ‘Candidatus Liberibacter solanacearum’. The role of carrot psyllid feeding and the psyllid‐associated bacterium in the damage formation are discussed.     

Differential belowground allelopathic effects of leaf and root of Mikania micrantha

This study evaluates the relative contributions of leaves and roots to the belowground allelopathic effects of Mikania micrantha. The hypothesis that leaves contribute more to the allelopathic effect than roots was experimentally tested. We assessed the allelopathic effects of aqueous extracts from Mikania leaves and roots on the seed germination and seedling growth of two co-occurring woody plants in southern China, Lagerstroemia indica L. and Robinia pseudoacacia L. The results showed that the aqueous extracts from Mikania leaves and roots had inhibitory effects on the woody species. Allelopathic activity depended on the concentration of the extracts, target species, and the extract sources (i.e., leaves vs. roots of Mikania). Leaf extract showed stronger allelopathic effects than root extract on germination percentage, initial germination time, speed of germination, and shoot height; while root extract had greater allelopathic effects on roots than leaf extract. The latter phenomenon might greatly promote the invasion success of Mikania due to more direct and effective allelopathy of root. Our results suggest that allelopathy of root extract on belowground biomass might be greater than that of leaf extract for some species in contrast allelopathy of leaf extract on belowground biomass might also be greater than that of root extract for other species, at least for their effects on root growth of the target species.     

IS POLYPLOIDY NECESSARY FOR TISSUE DIFFERENTIATION IN HIGHER PLANTS?

Measurements of relative DNA per nucleus of cells from various tissues show that cell differentiation can occur in the absence of polyploidy in higher plants. In Pisum polyploidy was present in roots, sepals, pods, pistils, and stamens but not in petals or leaves. In Triticum cells of leaves exhibited some polyploidy, but no polyploid cells were present in mature roots. No polyploid cells were found in any tissue of Helianthus examined (roots, cotyledons, stems, leaves, sepals, petals, pistils, and stamens). Therefore, as a general rule, polyploidy should not be considered essential in tissue or organ differentiation of higher plants. In Helianthus polyploidy is unnecessary for the completion of the life cycle.     

Soil-borne wheat mosaic virus inclusion bodies: structural, compositional and staining properties

Anatomy and cytochemistry of inclusion bodies induced by Soil‐borne wheat mosaic virus infection were studied in roots and leaves to learn more about the nature of inclusions and their roles in pathogenesis. Acid Fuchsin, Giemsa stain, Toluidine Blue and Trypan Blue stains facilitated visualization of inclusion bodies. Combined, simultaneous staining with Acid Fuchsin and Toluidine Blue clearly differentiated inclusion bodies from host nuclei. The overall anatomy, composition and structure of virus inclusions in leaves and roots were generally similar, as shown by phase contrast, differential interference contrast, epifluorescence, laser scanning confocal and transmission electron microscopy. Both were often closely associated with host nuclei; both were comprised of intertwined masses of tubular material, presumably endoplasmic reticulum, and in which varied numbers and sizes of vacuolar cavities occurred. Leaf inclusions, however, were typically larger and more vacuolate than those in roots. Lipids were found to be significant constituents of both the tubular and vacuolar components of inclusions, indicated by positive staining with Nile Red and Sudan Black. Inclusion bodies in both leaves and roots lost their structural and compositional integrity, eventually becoming disorganized and devoid of clearly identifiable components as host tissue aged and symptom expression advanced. Significant results of this study include the first published examination of virus inclusion bodies in root tissue, the degree of structural detail of inclusion body anatomy revealed by laser scanning confocal microscopy and the presence of an extensive lipid component in virus inclusion bodies.     

闽西南红壤侵蚀区黑莎草功能性状特征及其与土壤因子的关系

为了解黑莎草(Gahnia tristis)在南方红壤侵蚀区的适应状况,测定了长汀县红壤侵蚀区的黑莎草叶片、根系的功能性状及土壤理化性质,并应用数理统计方法分析了黑莎草叶片与根系功能性状之间的相关性,及其对土壤因子的响应。结果表明,黑莎草叶片表型性状在冬夏季间存在显著差异,叶长、叶宽、叶组织密度和叶绿素表现为夏季显著大于冬季,根系表型性状则更具稳定性,冬季的根系养分含量均高于夏季,养分的分配上叶片养分高于根系养分。叶组织密度与叶绿素含量呈显著正相关,与比叶面积呈显著负相关;根组织密度与比根长和比根面积均呈显著负相关,叶片和根系养分间均呈显著正相关,土壤碳、氮、磷含量是影响黑莎草功能性状主要因子。因此,黑莎草可通过调节功能性状以适应环境变化,可作为地带性植物应用于南方红壤侵蚀区的植被恢复和水土流失治理。     

Techniques for the regeneration and genetic transformation of Arabidopsis pumila: an ephemeral plant suitable for investigating the mechanisms for adaptation to desert environments

Arabidopsis pumila is a type of cruciferous ephemeral plant, which in China mainly grows in the desert environments of northern Xinjiang. A. pumila not only has a short growth duration, but also has high photosynthetic efficiency, seed yield, salt tolerance, and drought resistance. It is an ideal species for the study of environmental adaptations in ephemeral plants. We induced callus tissue formation on the roots and hypocotyls of 8-day-old seedlings, and on the leaves and petioles of 4-week-old seedlings, and obtained multiple adventitious shoots on these tissues grown on Murashige and Skoog induction medium supplemented with 0.5 mg/L 6-Benzylaminopurine and 0.1 mg/L α-Naphthalene acetic acid. Young roots, hypocotyls, leaves, and petioles could all induce calluses, but the induction rate was highest on young roots. In addition, the leaves and petioles of 4-week-old seedlings were used as explants, the Δ1-pyrroline-5-carboxylic acid synthase gene 1 of A. pumila controlled by 35S promoter of cauliflower mosaic virus was used as target gene, and hygromycin B was used as screening antibiotic to explore Agrobacterium tumefaciens GV3101 mediated transformation. The results showed that the callus induction rate of petiole explants was the highest when they were treated with Agrobacterium suspension (OD₆₀₀?=?0.6) for 10 min and thenco-cultured in dark for 2 days. The qRT-PCR results showed that the ApP5CS1.1 gene was overexpressed in the transgenic plants. These protocols provide working research methods for exploring the cellular level adaptative mechanisms of this species to desert environments.

     

大气增温对杉木幼树叶片及细根生理特征的影响

为揭示亚热带森林对未来全球变暖的生理响应特征,本研究以杉木为研究对象,利用开顶式增温方式模拟气候变暖,研究其对叶片和细根丙二醛含量、活性氧代谢、渗透调节物质含量以及抗氧化酶活性的影响。研究结果显示：（1）增温显著增加杉木叶片和细根的丙二醛含量,且叶片丙二醛含量显著高于细根,说明增温加剧了杉木叶片和细根氧化损伤,且叶片氧化损伤程度高于细根;（2）增温后,杉木叶片脯氨酸和可溶性蛋白含量降低,细根脯氨酸和可溶性蛋白含量则增加;（3）增温显著提高了杉木叶片过氧化物酶活性,对杉木细根抗氧化酶活性无显著影响;（4）增温后,杉木叶片和细根活性氧含量未发生显著变化,杉木叶片活性氧含量显著高于细根。综合分析表明,尽管增温增加了杉木叶片和细根的氧化损伤,但杉木可以通过提高抗氧化保护酶活性（叶片）和积累较多的渗透调节物质（细根）来维持体内活性氧代谢平衡。可见,杉木地上和地下部分器官间的相互合作与协调使杉木能有效地适应高温环境。