THE HORMONAL CONTROL OF ORGAN FORMATION IN CALLUS OF MEDICAGO SATIVA L. CULTURED IN VITRO

Organogenesis in alfalfa callus (Medicago sativa L. cv. ‘Regen S’) has been obtained by the transfer of callus from an induction medium containing growth regulators to a regeneration medium lacking growth regulators. The transfer of callus from induction medium containing high levels of 2,4-D and low levels of kinetin to regeneration medium resulted in the formation of shoots. Conversely, the transfer of callus from induction medium containing low levels of 2,4-D and high levels of kinetin resulted in the formation of roots. The pattern of organogenesis on regeneration medium was modified by the nutritional composition of that medium. When Blaydes medium supplemented with inositol and yeast extract was employed as regeneration medium, root organogenesis was inhibited. Root organogenesis was not inhibited by either Shenk and Hildebrandt medium or Gamborg's B5 medium. Shoot formation occurred on all of these media. A survey of the in vitro organ-forming capacity of 14 genotypic clones from the cv. ‘Regen S’ was conducted. The capacity to form organs differed quantitatively among the clones analyzed. A more detailed analysis of a highly responsive clone (RA3) and a poorly responsive clone (RA5) revealed no significant qualitative difference in their organogenic responses.     

STRUCTURE OF THE SHOOT APEX IN LACTUCA SATIVA

Histological observations of the leaf lettuce ‘Black Seeded Simpson’ showed the dormant embryo to possess two visible leaves and a flkat to slightly depressed plumular apex. Observations conducted over a 12-day period of germination and growth showed the development of L₁ and L₂, emergence of L₃ and L₄, and periodic changes in size of the apex which were associated with leaf emergence. Thus the dormant embryo of Lactuca appears to be considerably more advanced in development than was previously believed. The shoot apex appeared flat to slightly depressed at all developmental stages studied.     

LEAF HISTOGENESIS IN LACTUCA SATIVA WITH EMPHASIS UPON LATICIFER ONTOGENY

The leaf primordia of Lactuca sativa ‘Meikoningen’ develop from a subapical initial in the second layer of the tunica on the side of a fiat shoot apex. Subsequent growth of the subsurface lamina is initiated by submarginal initials which divide anticlinally to produce an adaxial layer and ***a biseriate abaxiallayer, and periclinally to produce a middle layer from which procambium differentiates. The protoderm is derived from the first tunica layer by continuous anticlinal divisions. The activity of the subapical and submarginal initials is completed when the leaf is 0.3 mm in length and 4.0 mm in width, respectively. Continued growth of the leaf to 130-150 mm results from intercalary cell division and enlargement. The mature venation is visibly delineated when the leaf is 25-30 mm in length. Laticifer and phloem cells are initiated by the same mother cells in the ***procambium. The former become non-septate laticifers by resorption of cross walls. They mature concurrently with the phloem and before the xylem.     

莴苣花药发育过程中钙的分布特征

减数分裂前,莴苣花药中的钙颗粒很少。减数分裂后,花药绒毡层细胞中的钙颗粒明显增加, 同时在花药药室基质中也出现许多细小的钙颗粒。刚从四分体中释放出的小孢子内钙颗粒很少,伴随着花粉外壁物质在小孢子表面的沉积,钙颗粒开始积累在花粉壁部位。随后,小孢子中开始出现钙颗粒。当小孢子开始形成液泡后,钙颗粒向其中聚集,伴随着小液泡融合成大液泡,体积较大的钙颗粒主要集中在液泡中,而细胞质基质中的钙颗粒很少。随着二胞花粉中的大液泡消失,花粉细胞质中的钙颗粒变得很少。在以后的发育中,只有花粉壁中积累较多的钙颗粒。在莴苣花药发育过程中,钙与绒毡层细胞的退化和小孢子液泡形成以及二胞花粉中大波泡的消失有关。而花粉外壁表面积累丰富的钙与以后花粉的萌发有关。     

印楝愈伤组织形成及其印楝素含量测定

取印楝（AzadirachtaindicaA．Juss）不同器官作外植体,研究培养基和继代次数对愈伤组织生长及其印楝素（Azadirachtin）生物合成的影响．印楝的不同器官（根、叶、茎及皮）均能诱导出愈伤组织,这些愈伤组织均有合成印楝素的能力．其中以叶诱导的愈伤组织生长速率及印楝素含量为最高．含有较低按盐的B5培养基有利于细胞生长,含有较高铵盐的MS培养基有利于印楝素积累,不含铰盐的White培养基对两者均不利．愈伤组织继代2—3代,有利于愈伤组织生长和印楝素合成。     

THE CHEMICAL COMPOSITION OF MYELIN IN ORGAN CULTURES OF RAT CEREBELLUM

Myelinating organ cultures of rat cerebellum were maintained in vitro for up to 130 days. Extensive myelination took place between 7 DIV (days in vitro) and 28 DIV. Centrifugation of a crude culture myelin fraction on a discontinuous gradient yielded three layers termed light myelin, heavy myelin and membrane fraction, which exhibited an ultrastructure virtually identical to that of comparable layers prepared from surviving littermates. However, culture myelin layers showed a gross deficiency of galactolipids with a relative increase in phospholipids. The 2,3′-cyclic nucleoside-monophosphate phosphodiesterase (CNP) activity was decreased in the culture myelin layers, but not to an extent comparable to the cerebroside deficiency. A form of “slow myelin maturation” takes place in vitro with both myelin cerebrosides and sulphatides increasing in cultures older than 60 DIV. The results indicate that CNS myelination comprises at least two phases, and that the second phase involving galactolipid enrichment of myelin can, under experimental conditions, be partly uncoupled from the first phase without affecting the morphology or ultrastructure of the sheaths.     

HISTOLOGICAL ANALYSIS OF ADVENTITIOUS BUD FORMATION IN CULTURED DOUGLAS FIR COTYLEDON

Initiation of adventitious bud formation in vitro from Douglas fir cotyledons required both cytokinin and auxin at concentrations of 5 μM BAP and 5 nM NAA. Histological observations showed that these adventitious buds arose de novo from cells residing in hypodermal layers. Development of adventitious buds in culture was characterized by the sequential appearance of four anatomically distinguishable structures: 1) meristemoid, 2) bud primordium, 3) shoot apex with needle primordia, and 4) adventitious bud. The anatomical structure of tissue culture-produced buds was similar to that of vegetative buds produced on intact plants. Cultured cotyledons capable of producing adventitious buds (bud culture) were compared with bud-callus and callus cultures initiated by 5 μM BAP plus 5 μM NAA and 5μM NAA alone without BAP, respectively. Results showed that, during early stages of the culture period (i.e., prior to the appearance of meristemoid structure), cell division of bud culture was mainly located in hypodermal layers, whereas for the other culture types, bud-callus and callus cultures, cell division occurred randomly in all tissues.     

莴苣卵细胞、合子与原胚细胞中钙的分布

用焦锑酸盐沉淀法对莴苣开花前后的卵细胞、合子与原胚细胞中的钙颗粒分布变化进行了观察。结果表明,开花前三天,刚形成的卵细胞内钙颗粒很少,开花前二天的卵细胞内钙颗粒开始增多,开花前一天的卵细胞形成了大液泡,建立了极性,细胞内的钙颗粒又减少。开花后、受精前的卵细胞的钙颗粒主要聚集在细胞核中。受精后合子中的钙颗粒又明显增多,在核质中分布一些较大的钙颗粒,在珠孔端大液泡中聚集了较多的絮状钙。二胞原胚中的钙颗粒又开始减少,多胞原胚细胞中的钙进一步减少,但原胚表面分布一层丰富的钙颗粒。探讨了钙在卵细胞分化成熟、受精以及原胚发育初期中的作用。     

INTERACTION BETWEEN HELIANGINE AND PYRIMIDINES IN ADVENTITIOUS ROOT FORMATION OF PHASEOLUS CUTTINGS

1. Heliangine at 110^–4 M promoted the adventitious rootformation in hypocotyls of cuttings taken from light-grown (1,900lux) Phaseolus mungo seedlings. The promotion was almost completelyreversed by 310^–4 M uracil, uridine, cytidine, oroticacid or 610^–4 M carbamoyl DL-aspartic acid, and partlyby 310^–4 M thymine or thymidine. Neither 310^–4M cytosine, adenine, adenosine, guanine, guanosine nor a combinationof 310^–4 M carbamoyl phosphate and 310^–4 M L-asparticacid reduced the promotion by heliangine.
2. Uracil did not reducethe inhibiting effect of heliangine onthe indoleacetic acidinduced elongation of etiolated Avenacoleoptile sections.
3. Helianginein an aqueous uracil solution was recovered unchangedafter24-hr incubation at room temperature.
4. The root formation ofPhaseolus cuttings was promoted also by2-thiouracil and 5-fluorouracil.The effect was reversed byorotic acid or carbamoyl asparticacid, but not by carbamoylphosphate plus aspartic acid.
5. Ribonucleaseat 100 µg/ml increased the number of rootsprotruded fromhypocotyls of cuttings by about 260%.
6. A possible interpretationfor the promotion of root formationby heliangine is offered.

¹ Contribution No. 15 from the Botanical Gardens, Faculty ofScience, University of Tokyo, Tokyo, Japan. ² Dedicated to Prof. Dr. H. SODING in commemoration of the 70thbirthday.     

莴苣及其变种果实的电镜观察

利用扫描电镜对安徽省栽培的莴苣Lactuca sativa及其变种的果实进行观察,结果发现涡阳县义门镇特产的苔菜既不同于莴笋L.sativa var.angustata,也不同于莴苣,应将其建立在莴苣种下作为新变种苔菜L.sativa L.var.yemensis X.S.Shen et R.L.Lu var.nov.更为合适。     

CHEMICAL CONTROL OF MORPHOGENESIS IN MOSS PROTONEMA

1. The protonema of the moss, Funaria hygrometrica, grows continuouslyin calcium-free liquid media.
2. The growth was promoted by additionof oxalate, although themorphogenesis resulting in formationof gametophytic buds onthe protonema was suppressed by theaddition.
3. Calcium oxalate promoted the growth of protonema,while at ahigh concentration (10^–2 M) it caused the formationofclumped protonema (falsebuds).
4. Addition of plant growthhormones, such as IAA, NAA, 2,4-D andgibberellin retarded thegrowth of protonema, while 2,4-D ata low concentration stimulatedthe growth of protonema.
5. Kinetin greatly stimulated the formationof gametophytic budsin the protonema, but these buds were foundto be morphologicallyand physiologically abnormal.

(Received January 29, 1965; )