日 蚕 雑 62(1) 85-87 (1993)

J. Seric. Sci. Jpn.

orted.

Growth of mulberry synthetic seeds

Materials and Methods
on vermiculite, sand and soil media
As described previously (Machii, 1992b), ad-

HIR0AKI MACHII1) ventitious buds were used as explants for the

and HIROAKI YAMANOUCHI production of synthetic seeds. These adventi-

tious buds were induced from immature leaves

of the mulberry variety 'Shin-ichinose' cultured

National Institute of Sericultural
on MS medium (Murashige and Skoog, 1962)
and Entomological Science
modified by the addition of 3% fructose and 5 tt
(Received August 4, 1992)
M benzyladenine, pH5.8, 1.0% agar. About 30

町 井 博 明 ・山 ノ内 宏 昭:ク ワ人 工 種 子 の バ ー ミ days after the start of the culture, adventitious

キ ュ ライ ト,砂,土 培 地 上 で の生 長 buds were excised from immature leaf cultures

and transferred to a 3% sodium alginate solu-

Recently, in many crops attempts have been tion. Then, the adventitious buds were dropped

made to obtain synthetic seeds using somatic into a 100mM calcium chloride solution one by

embryos produced by plant tissue culture. How- one for eucapsulation, resulting in the produc-

ever, these attempts have been successful in only tion of synthetic seeds. Immediately after this

several annual crops, such as alfalfa (Redenba- procedure, half of the synthetic seeds were sown

ugh et al., 1986, 1987), celery (Redenbaugh et al., on growth media, the remaining half being

1986; Sakamoto, 1990), lettuce (Sakamoto, 1990) stored in MS liquid medium containing 1.0mg/l

and carrot (Kitto and Janick, 1985a, 1985b). BA, 0.1mg/l NAA and 30g/l sucrose at 4•Ž for 30

On the other hand, in the case of mulberry days, and then sown on the growth media. The

tissue culture a method was developed to induce growth media consisted of vermiculite, sand and

adventitious buds directly from leaf cultures soil (clay loam) added to 100-ml Erlenmeyer

(Oka and Ohyama, 1981; Katagiri and Nishigu- flasks, which were all autoclaved at 121•Ž for 15

chi,1986). Moreover, it was demonstrated that a min. No nutrients and plant growth regulators

larger number of adventitious buds could be were added except for water. After sowing, the

induced more efficiently from immature leaves Erlenmeyer flasks were placed in a growth cabi-

isolated from winter buds than from mature net at 27°C with fluorescent lamps (60-80 u mol

leaves of shoots cultured in vitro (Saito and Kat- m-2 . s-1). After 30 days of incubation, germina-

agiri,1989; Machii, 1992a; Saito,1992). In a previ- tion, shoot growth and rooting were observed.

ous paper we reported that such adventitious As described in the previous paper (Machii, 1992

buds were useful materials for producing syn- b), when the expanded leaves emerged from the

thetic seeds of mulberry (Machii, 1992b). capsules, the growth stage of the synthetic seeds

In the present paper the growth and rooting of corresponded to germination. When the shoot

synthetic seeds of mulberry on three different stem emerged from the capsules, it was assumed

growth media, vermiculite, sand and soil are rep- that shoot growth had occured.

1-2 Ohwashi, Tsukuba, Ibaraki 305, Japan Results and Discussion

1) Present address: National Institute of Agro-

biological Resources, Kannondai, Tsukuba, Table 1 shows the growth of stored and non-

Ibaraki 305, Japan stored synthetic seeds on the different media.

-85-
 86 Journal of Sericultural Science of Japan Vol. 62 No. 1

Table 1. Growth of synthetic seeds on different growth media.

Mulberry variety: 'Shin-ichinose'.

The data were collected 30 days after sowing.

* Stored in MS liquid medium containing 1
.0mg/1 BA, 0.1mg/I NAA and 30g/l sucrose

at 4•Ž for 30 days before sowing on each growth medium.

media. The highest rates of germination, shoot

growth and rooting (75.0%, 62.5% and 25.0%,

respectively) were observed on the soil medium,

followed by the sand and the vermiculite media.

Fig. 1 shows the rooting of the synthetic seeds

grown on the three growth media, vermiculite,

sand and soil, respectively. Rooting was most

vigorous on the soil, followed by the vermiculite

and the sand media.

Thus, synthetic seeds which were sown on the

growth media without storage did not root on

Fig. 1. Synthetic seeds rooted on different
any of the growth media, whereas synthetic
growth media.
The growth media were 1; vermiculite, 2; seeds which were stored at 4•Ž for 30 days before

sand and 3; soil. The synthetic seeds were sowing rooted on all the growth media. In the

stored in MS liquid medium containing 1.0

previous paper (Machii, 1992b), synthetic seeds
mg/l BA, 0.1mg/l NAA and 30g/l sucrose at
did not root on the vermiculite medium, regard-
4•Ž for 30 days. The photo was taken 30 days

after sowing on the growth media. less of whether they were stored at 4•Ž or not

stored for some time before sowing. In this ex-

The non-stored synthetic seeds germinated on periment, synthetic seeds rooted on vermiculite

all the three growth media, with a rate of 18. 8% at the low rate described above. The finer size of

on vermiculite, 68.8% on sand and 43.8% on soil. vermiculite medium used in this experiment

However, neither shoots nor roots developed on than that described in the previous paper may

any growth media. On the other hand, more than account for the difference between the results in

50% of the synthetic seeds stored at 4•Ž for 30 the previous and present experiments. It is inter-

days after the procedure were able to germinate esting to note that 25% of the stored synthetic

and they developed shoots on all the growth seeds rooted on soil without the addition of nu-
 MACHII and YAMANOUCHI: Growth of synthetic seeds of mulberry 87

trients and plant growth regulators except for -282 .

water. KITTO, S. L. and JANICK, J. (1985b): Hardening
treatment increases survival of synthetically-
If the synthetic seeds produced by using so-
coated asexual embryos of carrrot. J. Amer.
matic embryos or adventitious buds could regen- Soc. Hort. Sci., 110, 283-286.
erate on soil, the impact on mulberry cultivation MACHII, H. (1992a): Organogenesis from imma-
for sericulture would be significant. Somatic ture leaf cultures in mulberry, Morus alba L.. J.
Seric. Sci. Jpn., 61, 512-519.
embryos are able to regenerate whole plants di-
MACHII, H. (1992 b): In vitro growth of en-
rectly on some media at one time. However, no capsulated adventitious buds in mulberry,
somatic embryos have been induced from mul- Morus alba L. Japan. J. Breeding, 42, 553-559.
berry tissue cultures so far, whereas whole MURASHIGE,T. and SKOoG, F. (1962): A revised
medium for rapid growth and bio assays with
plants are generally regenerated from adventi-
tobacco tissue cultures. Physiol. Plant., 15, 473-
tious buds through the formation of shoots follo- 497.
wed by that of the rooting system on different OKA, S. and OHYAMA,K (1981): In vitro initiation
media. It is thus interesting to note that 25% of of adventitious buds and its modification by
high concentration of benzyladenine in leaf
the synthetic seeds produced displayed a rooting
tissues of mulberry (Morus alba). Can. J. Bot.,
ability on soil in our material. 59, 68-74.
REDENBAUGH, K., PAA5CH,B. D., NICOL,J. W., KOs-
Acknowledgement sLER, M. E., VIss, P. R. and WALKER, K. A.
(1986): Somatic seeds: Encapsulation of asexu-
We are grateful to Dr. Katagiri for the critical
al plant embryos. Bio/Technology, 4, 797-801.
reading of the manuscript and to Ms. Hazeyama REDENBAUGH, K., SLADE,D., VIss, P. and FUJII, J. A.
and Ms. Umeda for their excellent technical as- (1987): Encapsulation of somatic embryos in
sistance. synthetic seed coats. HortScience, 22, 803-809.
SAITo, H. (1992): In vitro propagation of mulberry
References through adventitious-bud induction system. J.
Seric. Sci. Jpn., 61, 46-51.
KATAGIRI,K. and NIsmGucHl, T. (1986): Interspec- SAITO,H. and KATAGIRI,K. (1989): Adventitious
ific difference on adventitious shoot formation bud induction in leaves isolated from winter
in mulberry tissue cultures. J. Seric. Sci. Jpn., buds of mulberry. J. Seri. Sci. Jpn., 58,197-202.
55, 79-80. SAKAMOTO,Y. (1990): Development of synthetic
KITTo, S. L. and JANICK.J. (1985a): Production of seed system. International Bio Symposium '90,
synthetic seeds by encapsulating asexual emb- Nagoya, p. 218-224.
ryos of carrot. J. Amer. Soc. Hort. Sci., 110, 277