Dormancy in Laportea bulbils: experiments with buds cultured in vitro

Dormancy in bulbils of Laportea bulbifera was investigated usingbuds cultured in vitro. Buds excised from immature bulbils couldsprout in light, but scarcely in the dark. Regardless of lightconditions, buds isolated from mature bulbils failed to sproutwhile those from chilled mature bulbils could. GA and BA stimulatedthe sprouting of buds excised from dormant mature bulbils. Incontrast, ABA and morphactin inhibited the sprouting of budsexcised from nondormant chilled mature bulbils. These resultssuggest that, in dormant Laportea bulbils, the buds themselvesare in a dormant state which may be controlled by a balanceof activity between growth-inhibiting and -promoting substances. (Received October 29, 1976; )     

Understanding the germination of bulbils from an ecological perspective: a case study on Chinese yam (Dioscorea polystachya)

Background and Aims

Bulbils serve as a means of vegetative reproduction and of dispersal for many plants; this latter aspect making them analogous to seeds. However, germination of bulbils may differ considerably from seeds due to dissimilar anatomical structures and perhaps environmental cue perception. The few laboratory studies done on bulbils suggest that their germination is similar to that of seeds in the same habitats and to vegetative buds of winter-dormant plants. The present study is the first to examine how bulbil germination is controlled in nature in relation to dispersal (before vs. after winter of the same cohort) and to ambient temperatures.

Methods

Under laboratory conditions, temperature and light requirements for root and shoot emergences from bulbils of Dioscorea polystachya collected in September, 2005, February, 2006 (produced in 2005) and July, 2006 were determined. Effects of cold stratification and dry storage for releasing dormancy were tested on September and July bulbils. The phenology of dormancy release and of root and shoot emergences and the persistence of bulbils in soil were followed over time under field conditions.

Key Results

Although a low percentage of bulbils collected in July or in September produced roots, but no shoots, in the laboratory and field, these roots died within approx. 1 month. Regardless of collection date, cold stratification markedly increased root and shoot emergences. Bulbils sown outdoors in October produced roots and shoots the following March and April, respectively. The soil bulbil bank is short lived.

Conclusions

Bulbils of D. polystachya are similar to seeds of many temperate plants being mostly dormant when dispersed in summer or autumn and overcoming dormancy with cold stratification during winter. Adaptively, bulbil germination primarily occurs in spring at the beginning of a favourable period for survivorship and growth.     

Insurance against reproductive failure in a semelparous plant: bulbil formation in Agave macroacantha flowering stalks

Bulbils are small aerial rosettes that occur on the flowering stalks of semelparous Agave plants and in related families, and that are capable of acting as clones of the parent plant. We hypothesized that bulbil formation was inversely related to fruiting success in the flowering stalk, and we explored this hypothesis in A. macroacantha, a species from South-Central Mexico. Forty randomly chosen plants were divided amongst three treatments: (a) elimination of all floral buds, (b) exclusion of pollinators, and (c) control. We also studied 22 plants in which the flowering stalk had been felled by goat grazing. Between September and November 1991 we kept a record of the numbers of bulbils and capsules produced in each flowering stalk. Significant (P<0.0001) differences between treatments were found in the proportion of plants hearing capsules and bearing bulbils. The control treatment had the highest proportion of plants producing capsules, treatment a had the highest proportion of individuals bearing bulbils, while treatment b showed an intermediate response. In the goat-grazed group, 45% of the plants failed to produce any propagative structure after the stalk was cut, and half of all plants produced bulbils on the remaining stump. A significant inverse relationship between the numbers of capsules and the numbers of bulbils per plant was found for the three randomly assigned treatments. Our results suggest that once the production of the flowering stalk has been triggered and the death of the rosette is irreversible, bulbils may act as an insurance mechanism that increases the probability of successful reproduction of the genet.     

Physiological and Molecular Changes during Lily Underground Stem Axillary Bulbils Formation

Lily is regarded as one kind of the most economically important cut flowers in China. In this paper, lily cultivar ‘Aladdin’ (Lilium longiflorum × Lilium asiatic) was chosen as experimental material for its distinct high propagation coefficient in nature and three main conclusions were achieved. Firstly, our observation results revealed that the noticeable propagation coefficient of ‘Aladdin’ depended on strong capacity to form many underground stem axillary bulbils positioned on underground node of plant after 70 days of cultivation and having 1 cm flower buds. Secondly, in terms of physiological changes, zeatin riboside (ZR) relative concentration, soluble sugar and starch content peak when and where such bulbils are forming. Finally, two KNOX family Lilium genes were sequenced completely and one BELL family gene was sequenced partly; besides, there might be an interaction between KNOX-2 and BELL protein to accomplish their functions in bulbils organogenesis according to qRT-PCR results. As far as we know, it is the first time to study the formation mechanism of lily underground stem axillary bulbils. We illustrated physiological and molecular profiles, as well as offered a new prospect to lily molecular breeding in the future.     

淡黄花百合珠芽发育过程的形态学与解剖学研究

以野生淡黄花百合(Lilium sulphureum)为实验材料,通过形态学观察、石蜡切片方法对珠芽的发生规律、发育过程中的形态学变化及解剖结构特征进行了研究,以阐明淡黄花百合珠芽繁殖的生物学机制。结果显示:淡黄花百合的珠芽形成于地上茎叶腋处叶柄的近基部位置,由叶柄表皮以内数层薄壁细胞不断分裂和分化形成;珠芽的发育过程可划分为启动期、膨大期和成熟期三个时期。成熟珠芽外观上成圆形或椭圆形,珠芽中部的薄壁细胞含有大量淀粉颗粒。淡黄花百合的珠芽具有很高的萌发率(90%以上)。研究表明,珠芽营养繁殖是其生态适应的一种重要机制。     

The ecology of flower and bulbil production in Polygonum viviparum

Polygonum viviparum produces inflorescences which bear both sexual flowers and asexual bulbils in highly variable proportions. This paper investigates the proportion of flowers and bulbils per inflorescence at 12 sites in central Norway. It is shown that there is a 'trade—off between flowers and bulbils so that one cannot be increased without reducing the other'. On average, the balance is tilted in favour of bulbils, but there are significant differences in the proportion of bulbils per inflorescence among sites and among quadrats within 11 of the sites. Of these 11 sites there are 6 in which the proportion of bulbils per inflorescence is correlated with factors of the biotic environment. The factors include: (1) the amount of vascular plant cover, which is negatively correlated with the proportion of bulbils at 2 sites; (2) the cover of P. viviparum , which is positively correlated at 1 site; (3) the number of P. viviparum inflorescences, which is negatively correlated at 3 sites. The results suggest that both intra– and inter–specific factors of the biotic environment play a role in the control of bulbil and flower production in P. viviparum.     

Physiological activity in persistent bulbils of Agave vilmoriniana (Agavaceae)

Plants of Agave vilmoriniana are unique members of a reproductively diverse genus since they produce prolific numbers of both bulbils and viable seeds. Bulbils were observed to remain attached to the flowering stalk into a second summer, instead of abscising in the year they developed. Hence, a number of hypotheses were tested to establish the physiological basis for their longevity and evaluate their influence upon the plant's reproductive biology. Bulbils produced following anthesis were abundant (>2000 bulbils), large (0.44 g dry mass per individual), and their combined leaf surface area (one side) could exceed 3 m². Thus, the current-year inflorescence may act as a photosynthate source. Second-year, attached bulbils are healthy-looking, even after leaves of the parent plant have senesced, since maternal roots continue to absorb rain water. Rehydrated 2nd-yr bulbils resume Crassulacean acid metabolism, sometimes demonstrating net daytime tissue acidification. Water conservation by bulbils is very efficient, exhibiting very low cuticular conductances (0.016-0.0035 mm/s). Bulbils detach over all climatic seasons after having formed >3 and >18 root primordia in their 1st and 2nd year, respectively. About one-third of bulbils are capable of forming roots after a 1-yr storage at 35°C, when their water contents averaged 37%. Irrigated bulbils produce water-absorbing roots within 2 d, with de novo chlorophyll synthesis and nocturnal acidification commencing within 4 d. Two bulbils were observed to root and become established in shaded microhabitats during this 2-yr study of 29 plants.     

Dormancy in bulbils of several herbaceous plants: Effects of photoperiod,light, temperature,oxygen and gibberellic acid

Effects of some environmental conditions (photoperiod, white and colored lights, temperature, partial oxygen pressure) and growth regulators (gibberellic acid, 2-chloroethyltrimethylammonium chloride) on induction and release of dormancy of the bulbils ofDioscorea batalas, Laportea bulbifera, Elatostema involucratum andSedum bulbiferum were investigated. Bulbils were formed under short-day conditions inLaportea andElatostema, under long-day conditions inSedum, and irrespective of photoperiods inDioscorea. In all species exceptSedum, immature bulbils required light, particularly blue or far red, for sprouting (photo-sprouting stage), and mature bulbils required a cold treatment (thermo-sprouting stage). The duration of photo-sprouting and thermo-sprouting stages and the degree of dependency on light or low temperature of sprouting differed from species to species. Sprouting of chilled mature bulbils of these species was promoted by light, especially by red or green light. Both immature and mature bulbils ofSedum sprouted under short-day conditions. Continuous irradiation with blue, far-red and green light markedly inhibited their sprouting. Oxygen at high concentration inhibited the sprouting of immature bulbils inDioscorea; in the other species it promoted sprouting regardless of the maturation of the bulbils. Applications of gibberellic acid caused the sprouting of bulbils the absence of light, chilling or photoperiodic treatment in all species exceptDioscorea, in which gibberellic acid inhibited sprouting. Polyphenol oxidase activity was very high in the homogenates ofDioscorea bulbils, and increased further when the bulbils had been treated with gibberellic acid. In the other species, little or no such activity was observed.     

Meristem fate and bulbil formation in Titanotrichum (Gesneriaceae)

Titanotrichum oldhamii (a monotypic genus from Taiwan, Okinawa, and adjacent regions of China) has inflorescences bearing either showy yellow flowers or asexual bulbils. Asexual reproduction by bulbils is important in natural populations, and bulbil production increases in August and September at the end of the flowering season (which runs from June to the end of September). The bulbils are small (～1-2.5 mm long) and numerous. They consist of a small portion of stem (bract-stem) topped by opposite storage bracts that enclose a minute apical meristem. A secondary root develops from the side of the bract-stem. The floral meristem of T. oldhamii has three possible fates: (1) bulbil formation, (2) flower formation, or (3) bracteose proliferation. Bracteose proliferation rarely occurs and appears to be a developmental transition between the bulbiliferous and racemose inflorescence forms. It is strongly reminiscent of the floricaula and squamosa mutants of Antirrhinum. In the bulbiliferous form a single floral primordium, which would normally produce one flower, gives rise to ～50-70 bulbils by repeated subdivision of the meristem. This form of bulbil production appears to be unique to Titanotrichum. Occasionally a floral meristem divides, but the subdivision forms multiflowered units of up to four flowers rather than bulbils, suggesting that meristem fate is reversible up to the first or second meristem subdivision. In Titanotrichum, therefore, primordium fate is apparently not determined at inception but becomes irreversibly determined shortly after the appearance of developmental characteristics of the floral or bulbil pathway.     

Dormancy inDioscorea: Different temperature adaptation of seeds,bulbils and subterranean organs in relation to north-south distribution

Temperature dependencies of sprouting and germination were compared for subterranean perennating organs and seeds of ten closely related species of the genusDioscorea (Dioscoreaceae), a group of monocotyledonous summer perennials which are distributed from the tropics to the subarctic. The species used wereD. nipponica Makino,D. tokoro Makino,D. japonica Thunb.,D. tenuipes Franch. et Savat.,D. septemloba Thunb.,D. quinqueloba Thunb.,D. izuensis Akahori,D. bulbifera L. f.spontanea (Makino) Makino et Nemoto,D. pentaphylla. L. andD. alata L.; they are distributed from cold northern areas to warmer southern areas approximately in this order in and around Japan. Bulbil sprouting was also studied in those forming bulbils. Subterranean organs of the tropical species sprouted faster without any prior temperature treatment, whereas those of species from the more northern areas sprouted after prechilling. Northern species required longer, periods of prechilling for sprouting. On the other hand, with seeds or bulbils, the southern species required longer periods of prior temperature treatment for dormancy breaking. This difference in the length of dormant periods between seeds or bulbils and subterranean organs among the ten species may be related to their size and position of shedding; seeds or bulbils are small and are shed on the ground surface, whereas subterranean organs are large and are located below the surface. It is important to determine in other perennials whether the above relation between dormant features of seeds or bulbils and subterranean organs are common properties or not.     

Rapid clonal propagation of Pinellia ternata by tissue culture

Adventitious buds or protocorm-like bodies were regenerated directly from excised explants without intervening callus. Differences in the ability of regeneration were observed among different plant organs with bulbils showing the highest regenerative ability followed by leaf blade and petiole. Ability of vegetative propagation of bulbil could be maintained by alternate solid-and liquid-medium culture. Theoretically, 1.7×10²⁷ plantlets could be produced from a single bulbil by this technique within one year based on the production and rapid growth of protocorm-like bodies and adventitious buds. Concentration of MS salts, NAA and sucrose influenced not only root formation from the differentiated adventitious buds, but also root number and length. For root formation, the best combination was one-half strength MS salts with 3–5% sucrose and 1 mg/l NAA. The high survival rate of 96% was recorded when plantlets were transplanted into a mixture of vermiculite:loam soil:peat moss (1:2:1). Plants from in vitro culture were morphological similar to field-grown plants. The acute toxicity of crude extracts from protocorm-like bodies was about one-fourth that of extracts from tubers of field-grown plants when tested with white mice. Tissue culture has potential for clonal propagation of Pinellia ternata plants for commercial use.Abbreviations MS Murashige and Skoog (1962) - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA -naphthaleneacetic acid - BA 6-benzylaminopurine     

养分供应对3种舞花姜属植物繁殖方式的影响

该研究选用的3种舞花姜属植物中,毛舞花姜(Globba barthei)和双翅舞花姜(G. schomburgkii)自然条件下开花不结实,以珠芽进行繁殖;异果舞花姜(G. racemosa)则同时以种子和珠芽进行繁殖。在人工栽培条件下,高养分处理显著增加毛舞花姜的珠芽干重((1.11±0.10) g vs. (3.08±0.69) g,p=0.010 7)、珠芽数量((30.58±2.92) vs. (74.74±9.73),p=0.000 4)、珠芽大小((10.5±1.48) mg vs. (53.50±11.42) mg,p=0.001 5)、植株营养体重((3.99±0.23) g vs. (9.67±1.17) g,p=0.000 2)和双翅舞花姜的珠芽数量((51.74±3.64) vs. (108.71±21.52),p=0.018 1)。不同养分条件下两种舞花姜的开花数量无显著差异。人工去除珠芽、花对两种舞花姜植株当年生花和珠芽的数量无显著影响(p<0.05)。自然条件下异果舞花姜不同居群间珠芽大小、珠芽数、珠芽重、果实重、种子数和营养体重等均存在极显著差异(p<0.001)。异果舞花姜各居群植株珠芽数和营养体重(r=0.901,p<0.05)、珠芽重和珠芽大小 (r=0.849,p<0.05) 呈显著正相关;果实重和种子数(r=0.998,p<0.05) 呈显著正相关。土壤因子主成分分析表明前3个主成分提供的信息量分别为50.575%、18.204% 和11.883%,其中第一主成分中的全氮(0.959)、速效氮(0.885)和有机质(0.821)负荷量最大,第二主成分中土壤速效磷(0.824)负荷量最大。植株营养体重和土壤速效磷极显著正相关(r=0.906,p<0.01),珠芽大小与全氮(r=0.798)、速效氮(r=0.780)和有机质(r=0.821)均呈显著正相关(p<0.05)。果实重和种子数与土壤因子相关性不显著(p<0.05)。3种舞花姜的无性繁殖器官更易受到养分供应的影响,有性繁殖器官对养分供应变化响应不显著,有性和无性繁殖之间似不存在补偿关系。     

Minor differences with big consequences: Reproductive patterns in the genus Gagea (Liliaceae)

Reproductive patterns in ten species of Gagea Salisb. were compared by counts and measurements of bulbs, bulbils and flowers in large cohorts including all life stages. Two types of bulbils were found: taxa with “type I bulbils” start to develop a single to several bulbils as soon as the replacement bulb has reached a certain diameter and then continue to form them indefinitely throughout the life of the plant. “Type II bulbils” are only temporarily produced in immature, non-flowering plants of some species, but not in fully grown, flowering individuals, a phenomenon termed “reproductive switch”. Patterns of bulbil formation are species-specific: G. davlianidzeae, G. nigra, G. peduncularis, G. pratensis, and G. spathacea produce only type I bulbils; G. angelae, G. fedschenkoana and G. lutea develop only type II bulbils. Both bulbil types occur simultaneously in G. fragifera and G. villosa. The quantitative investigations demonstrate the existence of species-specific thresholds for the development of bulbils as well as flowers. Compared to the adult volume of the replacement bulb (where 90% of all plants flower), both types of bulbils have usually low thresholds: 0–5% (type I, all but one species) and 3–13% (type II). Inflorescences develop if plants attain between 38 and 60% of the adult bulb volume. Minor changes in patterns of bulbil formation and thresholds for their development may ensure survival of highly sterile taxa (e.g. G. spathacea, G. fragifera). This, in turn, can facilitate speciation in the genus driven by both hybridization and polyploidization.     

Loss of short-day requirement for dormancy breaking of bulbils from gibberellic acid-treated plants in Sedum bulbiferum

Sedum bulbiferum forms bulbils at superterranean nodes under long-day conditions, and the detached bulbils sprout after exposure to short-days [1]. When gibberellic acid was applied to the mother plant at the start of the long-day induction period, the number of bulbils formed increased slightly and these bulbils sprouted on incubation in the dark but not under short days of continuous light. However, when gibberellic acid was applied directly to detached bulbils during incubation, the short-day requirement for sprouting was conserved. Gibberellic acid application to the mother plants enhanced sprouting ability of detached bulbils when incubated under illumination with blue, green or far-red light. However, presence of gibberellic acid during bulbil exposure to light did not induced marked enhancement in sprouting under blue, green of far-red light. Thus, gibberellic acid application to the mother plant modified light and photoperiodic requirements for the sprouting of detached bulbils of S. bulbiferum.     

Studies on the growth and development of Oxalis latifolia

The growth and development of Oxalis latifolia was assessed on outdoor-grown potted plants raised from bulbs. Three main phases of growth appeared to be correlated with major physiological developments in the aerial and subterranean organs. Establishment (weeks 1 – 5) was characterised by a slow gain in plant dry weight accompanied by high relative growth rate (RGR) values for individual plant organs. The second phase of growth (weeks 6 – 12) was reproductive, as stolons grew and initiated bulbils at the apices. Bulbils had the capacity to produce their own shoot system and ultimately some primary bulbils supported secondary bulbils. Dry matter accumulation was in the order: bulbils > peduncles > leaves > petioles > stolons = roots > parent bulb. The RGR of individual plant organs declined together with the net assimilation (NAR) rates and the leaf area ratio of the shoots during the reproductive phase and beyond into the senescence phase (week 13 onwards). The results are discussed in relation to the problem of achieving weed control via cultural and herbicide treatments.     

Dormancy in Dioscorea: Sprouting promotion by inhibitors of protein synthesis in bulbils and rhizomes

The sprouting of immature and half-dormant mature bulbils andrhizomes in some species of Dioscorea was promoted by treatmentwith inhibitors of protein synthesis, although the sproutingof completely awake mature bulbils was inhibited. Gibberellicacid (GA₃) inhibition of the sprouting was relieved by cycloheximideand some base and amino acid analogues. Polyphenol oxidase activity in the bulbils decreased after incubationwith the protein-synthesis inhibitors. Possible involvement of protein synthesis in inducing dormancyof Dioscorea bulbils is discussed. (Received July 11, 1977; )     

A Landscape Perspective of the Stream Corridor Invasion and Habitat Characteristics of an Exotic (Dioscorea oppositifolia) in a Pristine Watershed in Illinois

The spatial distribution of exotics across riparian landscapes is not uniform, and research elaborating the environmental constraints and dispersal behavior that underlie these patterns of distribution is warranted. This study examined the spatial distribution, growth patterns, and habitat constraints of populations of the invasive Dioscorea oppositifolia in a forested stream corridor of a tributary of Drury Creek in Giant City State Park, IL. The distribution of D. oppositifolia was determined at the watershed scale mainly by floodplain structure and connectivity. Populations of D. oppositifolia were confined to the floodplain, with overbank flooding from the stream. Dioscorea oppositifolia probably originates in disturbed areas upstream of natural corridors, and subsequently, the species disperses downstream into pristine canyons or ravines via bulbils dispersing in the water. In Giant City State Park, populations of D. oppositifolia were distributed on the floodplain across broad gradients of soil texture, light, slope, and potential radiation. The study also examined the longevity of bulbils in various micro-environments to illuminate strategies for the management of the species in invaded watersheds. After 1 year, the highest percentages of bulbils were viable under leaves, and much lower percentages were viable over leaves, in soil, and in the creek (76.0±6.8, 21.2±9.6, 21.6±3.6, and 5.2±5.2%), respectively. This study suggests that management procedures that reduce leaf litter on the forest floor (e.g., prescribed burning) could reduce the number of bulbils of D. oppositifolia stored in the watershed.     

Bulbils contra seeds: reproductive investment in two species of Gagea (Liliaceae)

The reproductive biology of the sympatric species Gagea lutea and Gagea spathacea was analyzed morphologically and by resource allocation measurements. Both taxa reproduce vegetatively by bulbils. The hexaploid G. lutea regularly forms seeds, but flowering plants cease to produce bulbils. Seed set was never observed in nonaploid G. spathacea which does not stop vegetative reproduction when flowering. In this species, the pollen contains a high proportion of non-viable grains. Already sterile plants invest more resources (per cent of total nitrogen) into bulbils than G. lutea (10.9 vs. 5.9%). For flowering plants, the respective values are 6.1% (flowers) plus 18.4% (bulbils) for G. spathacea versus 14.8% (flowers only) for G. lutea. G. spathacea lost the ability to reproduce sexually and relies solely on vegetative reproduction. This seems to require the breakdown of the switch mechanism between the bulbil and flower formation (as in G. lutea) and a higher net investment in reproduction, hampering individual growth.     

Quantitative changes of batatasins and abscisic acid in relation to the development of dormancy in yam bulbils

As yam bulbils (Dioscorea batatas Decne.) grow on the motherplants, dormancy of the bulbils develops gradually. This isevidenced by the fact that the sproutability of bulbils decreasedin proportion to their sizes, 2–4 mm to 8–12 mmin diameter when bulbils are detached and sown in Petri dishes.This decreasing sproutability was seen until the stratificationat 5°C for dormancy release was completed. During the growth process of attached bulbils, the contentsof batatasins I and II increased markedly per surface area andper bulbil although on a fresh weight basis they remained constant.The increases of batatasin III and abscisic acid (ABA) weremore striking and were evident in all comparisons. These results corresponded to different localization of batatasinsand ABA in bulbil issues. All of 3 batatasins were localizedin the outermost layers of the bulbils, whereas ABA was distributedevenly throughout their tissues. These results suggest differentroles for batatasins and ABA in the regulation of yam bulbildormancy. ¹ This paper concerns K. H.'s Ph. D. dissertation presentedto Tohoku University based on work done at I. P. C. R. ² Present address: Biological Institute, Faculty of Science,Tohoku University, Sendai, Japan. (Received November 15, 1972; )