甜瓜子叶离体培养直接再生不定芽的形态学和解剖学观察

对甜瓜品种“西莫洛托”子叶离体培养直接再生不定芽过程进行了形态学和解剖学观察,结果表明：以子叶远轴面接触培养基培养能直接再生不定芽,以子叶近轴面接触培养基培养只能得到无序组织块;不定芽再生过程中,培养4d时,子叶变绿,但还不见有细胞分裂,约6d时,在子叶外植体的形态学下端切口处的近轴面到有局部的表皮细胞和亚表皮细胞分裂活跃,初步形成了拟分生组织,7－9d时,这些拟分生组织形成了肉眼可见的小突起,10－14d时,这些突起变得狭长,发育成幼叶或叶状体,它们在外植体上成族存在,但此时还没有典型的“芽”结构出现,15－20d时,成族突起形成幼叶丛,一些幼叶和叶状体的近轴面的基部出现叶腋分生组织,这些将外植体转入伸长培养基,3－5d后,幼叶充分展开,成丛叶状,多个叶腋分生组织同时发育成芽,再过2－7d,相继有不定芽从丛叶外植体上伸长,将伸长的不定芽切下,可促使外 植体上的其它不定芽伸长。     

Advances in Rabbit Embryo Culture during Organogenesis

Modified rodent embryo culture techniques were used to maintain 10 1/2 day rabbit embryos in vitro for up to 24 hr. In homologous, immediately centrifuged, heat inactivated serum containing 2 μCi/ml ³H-thymidine, observed status was consistently high at 12 hr but fell thereafter. Liquid scintillation counting and autoradiography revealed rapid uptake of radioactivity during the first 12 hr. The splanchnopleure of the inverted yolk-sac is only partially vascularised in the rabbit and is thus inadequate as a medium of metabolic exchange in extended culture. Nevertheless, this technique will enable a study of the morphology and biochemistry of teratology in vitro in a sensitive species.     

Direct Organogenesis in Hypocotyl Cultures of Tamarindus Indica

Direct differentiation of shoot buds from hypocotyl segments of 12-d-old seedlings of Tamarindus indica was obtained on Murashige and Skoog (MS) medium with or without growth regulators. The highest regeneration (66 %) and the maximum number of shoots (3 - 4) per explant were obtained from the explants on MS medium containing 6-benzylaminopurine (5 × 10-6 M). A maximum roots per shoot were produced on medium containing 3-indole butyric acid (5 × 10-6 M). The resulting plantlets were hardened and transferred to soil in pots where 75 % of them survived and resumed growth. Histological examination of explants suggests that the shoots were of de novo origin which would make this system suitable for transformation experiments. This revised version was published online in July 2006 with corrections to the Cover Date.     

Seed characteristics and testa textures of Pratensis, Orobon, Lathyrus, Orobastrum and Cicercula sections from Lathyrus (Fabaceae) in Turkey

The seed morphologies and testa textures of 23 taxa belonging to the Pratensis, Orobon, Lathyrus, Orobastrum and Cicercula sections of Lathyrus that can widely be found in Turkey were analysed. The findings obtained in this study and previous findings (34 taxa in total) were compared and interpreted at the level of the sections. Morphological properties including seed size, general shape, surface shape, colour, hilum length and width were measured under stereomicroscopy. The seeds were of spheroidal, subprolate and prolate (P/E = 0.90–1.58) types and medium in size. The smallest seeds belonged to Lathyrus inconspicuus var. inconspicuus (P = 2.19 ± 0.25 mm, E = 2.08 ± 0.16 mm) and the largest to L. sativus (P = 5.88 ± 0.74 mm, E = 5.36 ± 0.57 mm). The smallest hilum belonged to L. inconspicuus var. stenophyllus (0.38 ± 0.04 mm) and the largest to L. sylvestris (4.50 ± 0.58 mm). Testa textures such as papillae shape, dense, ribbing and presence or absence of a waxy layer were examined using scanning electron microscopy (SEM). In addition, some photographs included in this study were taken via stereomicroscopy and SEM.     

Electrophoretic Studies on Seed Protein of the Genus Lathyrus

SDS-polyacrylamide gel electrophoresis of total seed extracts revealed the presence of legumin-like polypeptides ranging in molecular weight from 42 kD to 89 kD in Lathyrus sativus and L. odoratus. The polypeptides of higher mol wt were however, absent in L. aphaca. Vicilin-like polypepides of mol wt 76, 54, 36, 33, 31, 20 and 17 kD were seen in L. sativus and L. odoratus and of mol wt 72, 58, 54, 52, 36, 33 and 20 kD in L. aphaca. Analysis of various seed protein fractions of L. sativus revealed the presence of a large number of albumin polypeptides varyingin mol wt range from 12.5 to 95 kD, when as glutelin (mol wt 18 to 80 kD) and prolamin (mol wt 25.5 and 26 kD) fraction polypeptides were relatively fewer. On the basis of similarities in seed polypeptide profiles, L. sativus and L. odoratus seem to be closely related, whereas L. aphaca appears to be distantly related.     

Direct Organogenesis from Mature Leaf and Petiole Explants of Eryngium Foetidum L.

Eryngium foetidum L. plants were regenerated from mature leaf and petiole explants through direct organogenesis without intervening callus phase. From leaf explants, adventitious multiple shoots raised on Murashige and Skoog (MS) medium supplemented with 4.43 M benzylaminopurine (BAP) and 0.57 M indole-3-acetic acid (IAA), whereas in petiole explants shoot regeneration occurred at 8.86 M BAP and 0.57 M IAAA. 80% of the leaf explants and 44% of petiole explants produced shoots after four weeks of culture. The regenerated plants were rooted on MS medium supplemented with 2.46 M indole-3-butyric acid and 2.88 M gibberellic acid. The plants were successfully established in the soil and showed 70.9% survival in the field.     

Direct Organogenesis from Petiole and Thin Cell Layer Explants in Sugar Beet Cultured In Vitro

Detrez, C., Tetu, T., Sangwan, R. S. and Sangwan-Norreel, B.S., 1988. Direct organogenesis from petiole and thin cell layerexplants in sugar beet cultured in vitro.—J. exp. Bot.39: 917–926. Plant regeneration was obtained by direct bud formation frompetiole as well as from thin cell layer explants taken fromsugar beet (Beta vulgaris L.) plants grown in vitro. The budswere mainly induced in the blade-petiole transition zone ofthe explants. High frequency bud regeneration was observed inpetiole and thin layer explants of 10 different breeding linesof sugar beet tested. Organogenesis resulted when petiole explantsexcised from 8-d-old seedlings grown on half-strength Murashigeand Skoog medium (MS) containing 3.0 mg dm^–3 naphthaleneacetic acid (NAA), 3.0 mg dm^–3 6-benzylaminopurine (BAP)and 1.0 mg dm^–3 2, 3, 5, triiodobenzoic acid (TIBA) werecultured on MS with 3.0 mg dm^–3 NAA and 3.0 mg dm^–3BAP. Thin cell layer strips isolated from shoot apices culturedon MS medium supplemented with 0–9 mg dm^–3 BAP or1.0 mg dm^–3 indolebutyric acid (IBA) formed adventitiousbuds on MS medium containing 0–5 mg dm^–3 NAA + 5.0mg dm^–3 BAP. Histological studies confirmed the sub-epidermalorigin of shoots. Key words: Beta vulgaris, direct organogenesis, in vitro culture, petiole, regeneration, thin cell layer     

大蒜花序轴离体培养器官发生途径的解剖学研究

以大蒜品种‘三月黄’(Allium sativum L.cv. Sanyuehuang)花序轴为外植体进行离体培养,对其器官发生过程进行了形态学和解剖学观察。结果显示:大蒜花序轴离体培养不经过愈伤组织,通过器官直接发生途径形成不定芽,其不定芽起源于大蒜花序轴维管组织韧皮部一侧周围的皮层薄壁细胞,属于外起源;皮层薄壁细胞经脱分化后,由最先形成的拟分生组织发育为茎尖分生组织,然后环绕其形成叶原基,茎尖和叶共同构成一个完整的不定芽;大蒜花序轴离体培养发生的不定芽与花苞中自然形成的营养芽发生部位一致。不定芽通过壮苗、生根培养可正常生根形成植株,如果继代培养周期超过21 d,鳞茎形成率可达90.56%。     

Direct Organogenesis and Plant Regeneration in Preconditioned Tissue Cultures of Lathyrus cicera L., L. ochrus (L.)DC. and L. sativus L.

This study demonstrates the importance of preconditioning ofsource tissue in regeneration of multiple shoot buds from severalspecies of Lathyrus. Preconditioned multiple shoots of Lathyruscicera L., L. ochrus (L.) DC. and L. sativus L. were obtainedby germinating seeds on Murashige and Skoog (MS) medium containing50 µM N⁵-benzylaminopurine (BAP) for 2 to 3 weeks. Multipleshoot bud formation occurred when epicotyl explants of preconditionedshoots were cultured on MS medium containing 5–50 µMBAP. No shoot regeneration was observed from epicotyl explantswhich were obtained from non-preconditioned shoots. Shoot budswere formed directly on explants without an intervening callusphase after 2 to 3 weeks of culture. Regenerated shoot budsformed healthy shoots which developed prolific and strong rootswhen transferred to MS medium supplemented with 2.5 µMnaphthaleneacetic acid (NAA). Lathyrus cicera L., L. ochrus (L.) DC., Ochrus Vetch, L. sativus L., Lathyrus pea, de novo differentiation, epicotyl, preconditioning with BAP     

Biochemical and Cytological Studies During Seed Development in Somaclones of Lathyrus sativus

Soluble proteins in six somaclones and the parent at three different stages of development of Lathyrus sativus seeds showed polypeptides of mol wts ranging from 20–165 kD on SDS-PAGE. Immature seeds showed eight distinct esterase bands and two peroxidase bands. Pattern for both the enzymes among somaclones was almost similar. During maturation a new isoform of peroxidase with high mobility appeared in all the somaclones, while one new isoform of intermediate mobility was visible in somaclones Bio L-08 and Bio I-08. Cytological analysis of all the somaclones confirmed the stability of the chromosome count of 2n=14. At metaphase I seven distinct bivalents and at anaphase I and II univalents showing complements of the seven chromosomes were visible. No structural or chromosomal aberration were observed among somaclones.     

In Vitro Clonal Propogation of Coleus forskohlii via Direct Shoot Organogenesis from Selected Leaf Explants

Present study provides an easy and efficient protocol for large scale clonal propagation of Coleus forskohlii, a threatened medicinal plant of commercial importance. Basal leaf lamina excised from upper three nodes of shoot was used as explant and its size, position, orientation and season of collection were initially optimized to select the most responsive explant condition. Enhanced shoot production and proliferation has been achieved on medium containing 2 μM BA + 0.1 μM NAA wherein, a highest number of 35 shoots/explant were produced. The regenerated shoots of varied length (3–5 cm) were transferred to root induction medium comprising of IBA, NAA and IAA (1–5 μM) in half-strength MS medium to determine the most suitable shoot length for proper root induction. Rooted plantlets were acclimatized in field conditions after proper hardening. Histological analysis was also carried out to confirm the nature of origin of shoot buds from leaf explants.     

Cyto-Histological Observations on Organogenesis of Rubus laciniatus Culture In Vitro

Leaf explants of Rubus laciniatus were cultured on Nitsch and Nitsch basic medium supplemented with either 6-BA 2 mg/L and NAA 0.1 kg/l or 2, 4-D 2.5 mg/l and NAA 0.1 mg/l. Adventitious buds were produced from leaf-like structures. Some ultrastructural observations of cells have shown that the most easy change part of a plastid during the cell differentiation in Rubus leaf explant culture in vitro was lamella system, storage starch and plastogluboli.     

油松成熟胚培养直接器官发生与植株再生

以油松（Pinus tabulaeformis Carr.）成熟合子胚为外植体进行了直接器官发生研究。在添加1~5 mg·L^-1 BA,或再配合添加1～10 mg·L^-1 2,4-D的DCR和MS培养基上均能诱导发生不定芽。在只添加1 mg·L^-1 BA的DCR培养基上不定芽诱导率最高,为90%。及时将不定芽继代于不含生长调节物质的DCR培养基上有利于芽的伸长。添加1 mg·L^-1 IBA的1/2DCR培养基最适于不定根的诱导,在该培养基上不定根诱导率达到47.1%。再生植株在蛭石基质上炼苗后转到温室培养,成活率高达86.4%,且生长正常。     

Organogenesis in vitro of tissues from mature conifers

Summary Shoot and embryolike structures were obtained in cultures of various tissues of mature conifer trees. Morphogenesis occurred in haploid and diploid tissues of female cones ofPinus mugo andPicea abies, although only rarely. Vegetative shoots dissected from early spring buds ofAbies balsamea, Picea glauca, andPseudotsuga menziesii were more responsive, with most of the organized growth occurring at the base of young needles. The following sequence of treatments was most effective: (a) collect twigs with buds shortly before budbreak, (b) cold store for at least 1 month, (c) force buds to break before excision of the vegetative shoots (keep submerged in water during excision), and (d) soak explant in malonic acid, 1 g/l, for 15 min before transfer of the shoots to the nutrient medium.     

Embryo-culture in Lathyrus

Development of 2-mm embryos of Lathyrus articulatus L. in vitrowas stimulated by placing them initially on an agar medium containing8 or 12 per cent sucrose. Germination took place after transferto a similar medium with 4 per cent sucrose. Successful cultureof 0.5-mm embryos was achieved under similar conditions butit was necessary to supplement the initial medium with 10 percent coconut milk. Using this technique, the hybrid L. clymenumL. x L. articulatus was raised from embryos which would haveotherwise aborted in vivo.     

Wheat Callus Culture: the Initiation, Growth and Organogenesis of Callus Derived from Various Explant Sources

Callus was obtained from mature excised embryos of wheat, fromnodal and internodal stem segments and from rachis segmentsusing the medium of Murashige and Skoog(1962)(M medium), containing1-0mg l^–1 2,4-D, and from immature embryos using the mediumof Green and Phillips (1975) containing 2 mg l^–1 2,4-D.Callus yield from mature embryos depended upon the cultivarused. No callus could be obtained from leaf segments. Callusderived from mature embryos and nodal stem segments was successfullymaintained by serial sub-culture on the M medium containing2,4-D for up to 3 years although its growth rate declined toa lower level as culture proceeded. Such cultures consistently produced roots when transferred toa medium containing a low level of 2,4-D or no 2,4-D. The presenceof the auxin was essential for continued proliferation of thecallus tissue. Shoot initiation was infrequent, did not occurafter the first few sub-cultures and could not be enhanced byvarious auxin and cytokinin additions to the medium. Callusderived from immature embryos did not have an enhanced potentialfor shoot initiation. Triticum aestivum, wheat, callus culture, organogenesis     

Organogenesis in Callus from Shoot Apices of Pisum sativum

Shoot formation was observed in callus from apical cells of pea (Pisum sativum L. cv. Century). Shoot apices from 4-day-old plants were macerated and the resulting cell masses grown on agar media. The callus formation and shoot production occurred within 4 to 6 weeks in defined media containing 0.2 to 5.0 μM benzyladenine and 1 μM naphthaleneacetic acid. While most callus produced one or more shoots at high frequency, root formation did not occur regularly. Plants obtained by these procedures were grown to maturity producing flowers and pods.