STUDIES IN BACTERIOSIS, XXVII FACTORS INFLUENCING INFECTION BY CORYNEBACTERIUM FASCIANS (TILFORD) DOWSON

Laboratory experiments showed that gram ( Cicer arietinum ) could be infected by Corynebacterium fascians. Parallel tests on sweet peas and gram confirmed the existence of host specificity among strains of the pathogen, and of differences in host susceptibility towards a given strain. Adventitious root formation appeared as a new, though rare, sign of infection. Inoculation of aerial dormant buds invariably caused infection whereas cotyledonary bud inoculation often failed. Sporing bacteria antagonistic to C. fascians in culture media were isolated from soil, but they failed to exert their influence in infection experiments in sand.     

THE ROLE OF CYTOKININS IN THE “FASCIATION” DISEASE CAUSED BY CORYNEBACTERIUM FASCIANS

The disease caused by Corynebacterium fascians can be imitated by treatment with kinetin. Cultures of the bacteria were grown on a purine-free medium of known composition. They require thiamine. Two assays for cytokinin were developed; one depends on the release of lateral pea buds from apical dominance and is highly specific but of only moderate sensitivity. The other depends on the retention of chlorophyll in senescing oat leaves and is very sensitive (detecting 2 × 10^–4 μg of kinetin equivalents), though it is somewhat less specific and, therefore, requires care in usage. Both give results in 3–4 days and are thus far more rapid than tissue culture methods. With these assays C. fascians has been shown to produce a chloroform-soluble cytokinin active in both tests. The substance is stable to heat in acid or basic media, is soluble in non-polar solvents, and behaves as a base. It is precipitated from water solution by Ag ions and may, therefore, be a purine derivative. Pea tissue infected with C. fascians, but not uninfected tissue, yields a compound of similar solubilities and biological activity. Reasons are given for believing that the synthesis of cytokinins may be important, not only for C. fascians, but also for many other plant parasites.     

COMPARATIVE ANATOMY OF THE SEED COATS OF GNETUM AND THEIR PROBABLE EVOLUTION

The seed coats of Gnetum gnemon L., G. ula Brongn., G. montanum f. parvifolium (Warb.) Mgf. and G. neglectum Bl. consist of three layers. The outer layer or sarcotesta is mostly parenchyma but contains some sclereids and fibers and a series of simple vascular bundles. The middle sclerotesta forms masses of sclereids in varying shapes and numbers, sometimes extending as a basal plate, and is usually thicker near the micropylar tube. The second layer also contains a series of small vascular bundles that reach the apex. Depending on the species, the middle layer is sometimes nearly free from the outer layer, may be partially fused with it, or completely fused to it at maturity. The innermost layer of the seed coat constitutes the endotesta which is membranous and only rarely contains sclerenchyma. It possesses a dichotomous venation system with varying degrees of anastomosing, depending upon the species. The above species show qualitative and quantitative differences in their sclerenchyma and laticifers. Seed coat anatomy may be useful in the diagnosis of some species. The trends of evolution of seed coat structure within these four species of Gnetum are discussed, and a comparison of tissue layers and vasculature with certain fossil pteridosperms is made, especially in the Trigonocarpales     

GIBBERELLIN-LIKE SUBSTANCES OCCURRING IN THE SEED OF PHARBITIS NIL CHOIS. AND THEIR CHANGE IN CONTENTS DURING THE SEED DEVELOPMENT

The amount of gibberellin-like substances in the seed of Pharbitisnil increased in parallel with the growth of the seed, and attained20 days after anthesis to a maximum of 0.115 µg gibberellinA₃ eqivalent per seed, when the seed reached its maximum freshweight or four-fifths of its final dry weight. At this maximumlevel, 0.03 µg and 0.1 µg gibberellin A₃ equivalentswere localized in the embryo and in the "endosperm", respectively.Three gibberellin-like substances (Factors I, II and III) wereseparated upon paper chromatography. In view of the changesin amount of the factors with respect to the seed maturation,these factors, especially Factor II, in the "endosperm" andembryo were assumed to participate in the initial or the mainpart of growth of the embryo. Dwarf rice seedling and maizedwarfs 1, 3 and 5 responded to the three factors nearly in thesame way. Pharbitis dwarf, however, responded only to FactorI, but not to Factors II and III. (Received February 21, 1963; )     

中国水鳖科植物种皮微形态特征及其系统学意义

通过扫描电镜对国产水鳖科植物(包括6属13种)的种皮微形态特征进行观察,并作了系统描述。根据种皮细胞形态、外种皮表面纹饰和内种皮内层小瘤状突起的特点将水鳖科植物的种皮微形态特征划分为3种类型,即海菜花型(海菜花属)、水鳖型(水鳖属)和苦草型(苦草属、水筛属、虾子草属和黑藻属),并作出了分属检索表。本文结果表明,种皮微形态特征可作为该科族、属以及属内种级水平分类的依据,对探讨属间关系和该科的系统发育关系亦具有重要的价值。种皮微形态特征支持Hutchinson(1959)和Eckhardt(1964)将海菜花属和水鳖属分别作为一个独立的族处理的观点。苦草属、水筛属和虾子草属种皮微形态特征的高度相似性表明它们间有密切的联系,不支持将它们置于不同亚科和族的分类处理。黑藻属虽与上述3属近缘,但其外种皮特征则较为独特,因此与水筛属放在不同族中更为合理。本文种皮微形态特征的研究结果支持iki1937)和Shaffer-Fehre(1991b)等关于水鳖科与茨藻科近缘的观点。     

中国茨藻科植物种皮微形态特征及其系统学意义

本文应用扫描电镜对国产茨藻科植物的外种皮和内种皮微形态特征进行了研究,结果表明茨藻科植物在种子表面细胞形态、表面纹饰和内种皮内层细胞形态及小瘤状突起的大小和密度等方面存在差异,可作为种级水平分类的依据,并首次发现外种皮微形态特征也与内种皮内层小瘤状突起一样具有重要的系统学意义。根据种皮微形态特征的研究结果,我们支持Miki(1937)和Shaffer-Fehre(1991b)等关于茨藻科与水鳖科近缘,而不应放在眼子菜目的观点。对弯果茨藻Najas ancistrocarpa A．Br．ex Magnus和草茨藻N.graminea Del.外种皮的研究结果表明其外种皮细胞表面具瘤状颗粒而非网状加厚。     

THE ROLE OF POPULATIONAL DIFFERENTIATION IN EXPERIMENTAL INFECTION OF PROSOPIS BY PHORADENDRON

Experimental infection of Prosopis species with the mistletoe Phoradendron tomeniosum subsp. tomentosum in a uniform garden demonstrated that differences in infection success between species, populations, and individuals can be caused by genetically-based differences in both hosts and parasites. Prosopis glandulosa var. glandulosa was least resistant to infection; P. laevigata was more resistant; and P. torreyana was most resistant. A population of P. glandulosa var. glandulosa grown from seeds collected in the lower Rio Grande valley was significantly more resistant to infection than three populations of the same variety from central and northern Texas. The higher resistance of the Rio Grande valley population appears to be due to introgression with P. laevigata. The experimental population of Phoradendron tomentosum subsp. tomentosum from northeastern Mexico and the population of the same subspecies from southern Texas had greater infectivity than did those from central and northern Texas, corresponding to the greater resistance of the southern Prosopis populations.     

THE ROLE OF THE CONNECTIVE TISSUE GROUND SUBSTANCES (MUCOPOLYSACCHARIDES) IN ALLERGIC INJURY

The basic histologic reactions of the classic allergic diseases and of several systemic diseases in which allergic mechanisms appear to operate are described and illustrated. Particular attention is drawn to the ground substances—mucopolysaccharides—which constitute important elements of connective tissue and vascular structure. The intimate locus of the allergic reaction appears to be in and to involve a swelling of such substances. It is suggested that antibodies (and possibly antigens) may be attached to these mucinous ground substances of the connective tissues.     

CHANGES IN THE CONTENT OF GIBBERELLIN-LIKE SUBSTANCES IN RIPENING SEED AND POD OF LUPINUS LUTEUS

The amount of gibberellin-like substances in seed, pod, embryoand "endosperm" of Lupinus luteus in relation to their developmentwas studied by means of the rice seedling test. The amount of gibberellin-like substances per seed increasedremarkably at the first stage of development, when the growthof seed and pod was very slow, and attained its maximum at suchan early stage as the dry weight of seed reached a value ofonly 10 per cent and that of pod 28 per cent of their respectivemaximum levels. Changes in amount of gibberellin-like substancespresent in the pod were very closely related to the changesin growth of pod. Gibberellin-like substances equivalent to0.3 µg gibberellin A₃ per seed and 0.028 µg perpod were found at the 20th day after anthesis, and no activityin mature seed and 0.09 µg per pod at the 50th day. Noactivity was found in the embryo itself at all stages, indicatingthat the gibberellin-like substances in seed occur only in the"endosperm," i.e. in the tissues that will eventually form testain fully ripened stage. (Received December 18, 1962; )     

国产酸浆属（茄科）的叶表皮和种皮特征及其分类学意义

在光学显微镜和扫描电镜下观察了茄科酸浆属５种２变种及邻近属２种植物：辣椒和龙珠的叶表皮及种皮特征,发现上述植物的叶上、下表皮细胞表面观形状为不规则形,垂周壁浅波状、波状或深波状;一些类群的气孔器在上、下表皮均存在,另一些类群的气孔器则仅在下表皮存在,其类型一致为无规则型。扫描电镜下叶下表皮的特征,包括角质膜以及气孔外拱盖和拱盖内缘的特征,有一定的分类学价值。此外,种皮纹饰可作为区分酸浆属、辣椒和龙     

THE ANTIBACTERIAL SUBSTANCES OF THE MARINE ALGA STICHOCHRYSIS IMMOBILIS (CHRYSOPHYTA)12

Two substances with antibacterial activity have been concentrated from the culture medium of Stichochrysis immobilis before senescence occurred, and purified, by a Sephadex G-25 filtration followed by 2-dimensional thin-layer chromatography on Merck F-254 silica gel plates. One of the substances is obviously a peptide; 11 different amino acids are linked in a 30 amino acid molecule (MW 3400). It was not possible to assign a chemical group to the second substance (MW ? 2600); it may be related to “humic acids” (Kalle's gelbstoff). Growth of the most sensitive bacteria was completely inhibited by 20-fold, or higher, concentrations of substances from the algal culture filtrate (assuming no loss of substances during the concentration processes). The 54 strains of test bacteria were of various origins, though mostly isolated from algal cultures or obtained directly from the marine environment.     

VASCULAR INFECTION OF COTTON BY XANTHOMONAS MALVACEARUM (E. F. SMITH) DOWSON

Cotton seedlings raised under glass from seed pre-soaked in a water suspension of Xanthomonas malvacearum , and watered by sub-irrigation only, developed atypical dull green flaccid areas extending from the periphery of the cotyledons. Later symptoms that variably developed were: vascular discoloration, not necessarily continuous, in the petioles of affected cotyledons, in the hypocotyl, and in parts of the plant above the cotyledonary node; premature withering of cotyledons; dull green flaccid areas in true leaves, not necessarily the lowest, later turning brown and drying, with usually a chlorotic margin; parenchymatous attack in stems, petioles and leaves. Evidence is adduced, from the progression of symptoms and from associations between them, and from isolations of the pathogen from various parts of the plant at different stages, that the course of this type of infection is primarily vascular, progressing from the edges of cotyledons into the hypocotyl and thence upwards in vascular tissues. Thence the infection may break out at any time into parenchymatous tissues, leading to the more characteristic forms of the disease, of greater or less severity according to the susceptibility of the parenchyma at the time of its invasion. Similar symptoms have been observed in field plantings, and it is suggested that vascular infection is of greater and wider significance in the epidemiology of bacterial blight than has hitherto been commonly accepted.     

PECTIC ENZYMES SECRETED BY VERTICILLIUM DAHLIAE AND THEIR ROLE IN THE DEVELOPMENT OF THE WILT DISEASE OF COTTON

An isolate of Verticillium dahliae obtained from Uganda was highly virulent to young cotton plants under greenhouse conditions. A hyaline variant which often appeared in culture was as virulent as the parent isolate, but preliminary experiments indicated that it did not survive as long in unsterile soil. The parent isolate grew rapidly in cotton plants after root inoculation and was isolated from stems and leaves well before the appearance of disease symptoms visible to the naked eye.
Protopectinase was produced in the absence of pectic materials, but more active preparations were obtained when media contained pectic substances. In general, there was a close correspondence between the protopectinase activity of culture filtrates and the toxicity of these filtrates to parenchymatous cells. Some separation of the two activities was obtained by heating enzyme solutions or by plasmolysing the test tissue.
Protopectinase solutions had little pectinesterase activity but rapidly reduced the viscosity of solutions of pectic substances. In general, the properties of protopectinase and the viscosity-reducing enzyme were similar.
Young cotton shoots wilted rapidly when placed in cell-free filtrates from cultures of the pathogen. Wilting was delayed under conditions unfavourable for transpiration. Evidence was obtained which showed that wilting was caused by the uptake of thermostable compounds of high molecular weight which impeded the upward flow of the vascular sap. Pronounced vascular browning was obtained only when solutions containing protopectinase were used. Wilting and vascular browning were obtained with solutions having little pectinesterase activity; in contrast, a solution having high pectinesterase activity produced relatively little vascular browning.     

ON THE DEVELOPMENT OF MACROSCLEREIDS IN SEED COATS OF PISUM SATIVUM L.

Macrosclereid differentiation was investigated by light and electron microscopy in pea testae during the transformation of protodermal precursors to the mature sclereids. The protodermal cells divide anticlinally and elongate into the macrosclereid layer during seed coat development. Young sclereids have elongate nuclei, plastids become somewhat granal during cellular maturation, vacuolation appears to be an autolytic process, and the cells have dense arrays of endoplasmic reticulum and ribosomes. Considerable dictyosome activity and microtubule development is observed as the secondary wall is produced. Many coated vesicles are associated with and fuse with the plasmalemma. During development, the outer tangential wall area of the macrosclereids acquires a definite cuticle and subcuticular layer. Also, at this time the sclereid walls under the subcuticular layer display semicircular microfibril orientation. The sclereid walls adjacent to the hypodermis become multilayered. As the macrosclereids near maturity, the “light line” becomes discernable in the light microscope at the junction of the cellulosic tips of the macrosclereids and the subcuticular layer. This “light line” is prominent using interference optics and is an osmiophilic layer in the electron microscope. This layer may represent the suberin “caps” reported by earlier workers.     

PRODUCTION OF HYALINE HAIRS BY INTERTIDAL SPECIES OF FUCUS (FUCALES) AND THEIR ROLE IN PHOSPHATE UPTAKE1

A field study to determine the precise times of year at which three intertidal species of Fucus start to produce hyaline hairs and cease producing such hairs was conducted on the Isle of Man, U.K. Hairs were first observed during February, and within 6 days of their initial appearance, all tagged plants of all species at all tidal heights on the shore possessed hairs. Hair production continued until the beginning of October, at which time Fucus plants growing at the lowest stations (+ 3.0 m) had glabrous apical growth. Hair production continued later into the year for plants growing higher on the shore, and it was not until mid-November that glabrous apical growth was observed in all plants. Phosphate uptake rates of pilose (hairy) and glabrous (hairless) apical sections were measured in November 1988 for F. spiralis L. and in January 1989 for F. spiralis and F. serratus L., at phosphate concentrations ranging from 0.8 μM (ambient seawater) to 9.0 μM. In ambient seawater, pilose plants of F. spiralis removed phosphate 2–3 times faster than glabrous plants, whereas the uptake rates of pilose plants of F. serratus were about 50% greater than those of glabrous plants. The differences between uptake rates of pilose and glabrous plants of both species were smaller or nonsignificant at higher phosphate concentrations. The field and laboratory data are consistent with the hypothesis that hairs are formed in Fucus as a response to increased nutrient demand and that hairs facilitate the uptake of nutrients from seawater at concentrations typical of natural situations.     

蚂蚁对澜沧舞花姜种子散布及种苗空间分布格局的影响

 在野外系统观测了澜沧舞花姜（Globba lancangensis）18个果实共216粒种子的散布过程。共有10种蚂蚁参与了澜沧舞花姜的种子散布,距离为0.01～3.35 m,平均距离(0.47±0.03) m（平均值±SE,n=216）。其中最重要的３种蚂蚁是横纹齿猛蚁（Odontoponera transversa）、大头蚁(Pheidole sp.)和黄足厚结猛蚁(Pachycondyla luteipes),其出现频率分别为61%、50%和28%,散布的平均距离分别为(0.60±0.09) m、(0.20±0.01) m和(0.32±0.05) m。从总体上看,蚂蚁促进了种子的分散,降低了种子的聚集程度。横纹齿猛蚁对于种子上的油质体最为敏感,对人工去除了油质体的种子不搬运,对种子散布距离较远,暗示了其与澜沧舞花姜之间可能存在更紧密的互惠关系。野外样方调查结果表明,在3种舞花姜属植物中,以种子繁殖为主的澜沧舞花姜种苗之间的平均最近距离为(36.8±1.45) cm（平均值±SE,n=74）,显著大于以珠芽繁殖为主的毛舞花姜（Globba barthiri）的(29.8±2.70) m（n=34）（t73,33=2.11,p=0.037）和异果舞花姜（Globba racemosa）的(28.7±3.16) cm（n=32）（t73,31=2.33,p=0.022）;澜沧舞花姜的种苗聚集程度（Z=-1.70±0.19）显著小于毛舞花姜（Z=-2.58±0.37,t73,33=2.36,p=0.020）和异果舞花姜（Z=-3.28±0.53）（t73,31=3.54,p=0.001）。这说明相对于毛舞花姜和异果舞花姜,蚂蚁对种子的散布作用显著增加了澜沧舞花姜种苗间的平均最近距离,降低了居群的聚集度。     

苏芸金杆菌伴孢晶体和芽孢孢衣的蛋白质组分及其与毒力的关系

研究了苏芸金杆菌(Bacillus thuringiensis)3个变种6个菌株的提纯伴孢晶体和芽孢对大蜡螟(Galleria mellonella)和大菜粉蝶(Pieris brassicae)的毒力、晶体的蛋白质和多肽成分及芽孢衣中的类晶体蛋白质成分.生物测定表明,晶体毒力高于芽孢,在总数量相同的情况下,晶体和芽孢近1:1的混合物的毒力高于单独的晶体或芽孢.芽孢衣中存在一种类似晶体蛋白质的成分,无晶体突变株及无效野生株的芽孢则无此种蛋白质且对两种昆虫无毒.变种wuhanensis和变种galleriae的晶体含MW138000的主要蛋白质和63000的次要蛋白质,经碱性缓冲液溶解后,上清液含MW138000的蛋白质,沉淀中含MW63000的蛋白质;变种aizawai的晶体中仅含138000的蛋白质.对大蜡螟无毒的HD-11(var.aizawai)晶体的蛋白质成分有别于上述晶体,其胰蛋白酶消化物的SDS凝胶电泳图型显示少2条多肽带,但对大菜粉蝶仍有效.结果表明,苏芸金杆菌的芽孢在昆虫病理中有重要作用,伴孢晶体的蛋白质和多肽成分与它们对昆虫的毒力特性之间有密切关系.