新テッポウユリとトランペットハイブリッドの遠縁交雑ユリの作出

誌名誌名 香川大学農学部学術報告

ISSNISSN 03685128

巻/号巻/号 65

掲載ページ掲載ページ p. 11-19

発行年月発行年月 2013年2月

農林水産省 農林水産技術会議事務局筑波産学連携支援センターTsukuba Business-Academia Cooperation Support Center, Agriculture, Forestry and Fisheries Research CouncilSecretariat

Tech. Bull. Fac. Agr. Kagawa Univ.， Vol. 65， 11 ~ 19， 2013

Production of Interspecific Hybrids between Lilium x戸rmolongiand Trumpet Hybrid Lilies

Seiichi Fukai， Jitlam Natenapitラ AsamiMochizuki， A卯 miHanazono and Takako Narumi

Abstract

Interspecific hybridization between L. x戸rmolongiand Trumpet hybrid lilies was done via cut style pollination

followed by ovary-ovule culture. All tested cross combinations produced progenies， but germination frequencies of

cultured ovules varied depending on th巴pollenparents and crossing year. The obtained LT progenies showed irト

termediate morphology with low pollen fertility. High-temperature treatments restored the pollen fertility of th巴 LT

hybrids. The pollen was backcrossed with L. x formolongi yielding triploid progenies. Genomic in situ hybridization

(GISH) revealed that the triploid lilies had LLT genomic constitution with some homoeologous recombinar山， indi-

cating that the fertility restored pollen is an unreduced gamete.

Keyword : GISH， Lilium， LT， ovarγ-ovule culture

Introduction

Lilies， on the world cut flower market， represent a rapidly

growing sector over the last several decades. The commercial

success of that sector can be continued through the progress of

lily breeding. Conventional crossing techniques have expand回

ed the breeding possibilities through interspecific hybridiza-

tion facilitated by cut欄 stylepollination， followed by embryo

rescue or ovary-ovule culture (1. 21. U sing these methods， wide

variations in flower color and shape have been produced.

Important issue of propagation science in lily is no longer the

development of methods for increase in number like micro-

propagatlOnヲ butrather understanding the reproduction system

that links both seed production and creation of variation in

plants. Better understanding of reproductive propagation with

the combination of different genomes in lily is useful in prac-

tical horticulture and also brings insight into the reproduction

system.

Lilium X formolongi hort. is a hybrid of L. longiflorum Thunb

(L) and L. formosanum. A. Wallace. In fact， L. X戸rmolo昭 i

is the only L hybrid lily for a cut flower that can be propa-

gated by seeds. Considerable improvements in flower shape

(up-right floret) and flowering characteristics (巴arlyor late

flowering) have been achieved. Nevertheless， L. X formolongi

produces only white flowers. It would be interesting to estab-

lish a seed-propagated L lily having a colorful flower.

Aurelian hybrids， colorful garden lilies， have been bred

from some trumpet lilies (T) including L. centifolium Stapf

ex Elwes， L. henryi Baker， L. regale E. H. Wilson， and L. sar-

gentiae E. H. Wilson. The hybrids show color白1flowers with

vigorous growth and disease resistance. Attempts have also

been made to introduce characteristics of L. regale into some

Oriental (0) species (3. 41. To introduce yellow flowers into 0

hybrids， T hybrids have been used¥O). Although T hybrids are

thought to be useful as breeding material to produce color

trumpet liliesへcrossesbetween L and T have not been stud-

ied intensively. Fukai and Tsuji 171 reported that progenies be-

tween L. x formolongi and white flower Asian trumpet lilies

showed early flowering. The combination of early flowering

and seed-propagation ability in L. X formolongi and color白l

petal color and disease resistance in Aurelian hybrids is an

interesting challenge to obtain new type lilies.

The int巴rspecifichybrids 0白enshow sterility. Restor:ation

of pollen fertility is necessary for use of the hybrids in fur-

ther breeding programs or propagation. Somatic chromosome

doubling using some chemicals such as colchicine or oryzalin

has been applied to achieve pollen germination 18.91. However，

severe limitation of fertile pollen produced by somatic chro-

mosome doubling has been recognized as lack of recombina-

tion. The us巳 ofsexual po叶lyploi凶di包za抗tωion(ωun町1

can oveぽrc∞omet白hiおslimi江ta低tiぬon正1ピ110叫1.However， the main problem is

that production of an unreduced gamete is genotype“specific

and lInstableflu-Recently，NatcnaPitdol.(12)reported that

high-temperature treatment of flower buds undergoing meiosis

can restore pollen germination ability in fertile interspecific

hybrid lilies.

This paper describes the performance of L-T crosses and

backcrosses of the hybrids through restoration of pollen fertil-

12 Tech. Bull. Fac. Agr. Kagawa Univ.， Vol. 65. 2013

ity using high-temperature treatment. To evaluate the intro-

gression of different genomesラ thechromosome composition

of backcross1 (BC1) was analyzed using genomic in situ

hybridization (GISH).

Materials and Methods

Plant materials

Seeds of L. X formolongi‘Augusta' and ‘White Lancer'

were obtained from commercial sources. The seeds were sown

in November in a greer由ousewith minimum temperature main-

1ained at 120

C. Seedlings with 2-3 leaves were transferred to

an unheated gre巴nhouse，where they were grown until flower.司

mg.

Bulbs of Aurelian hybrids and species (Tables 1， 2) were

obtained from commercial sources or breeders: All bulbs were

grown in the greenhouse without heating. Harvested pollen of

each genotype was kept at 40

C until further use.

Pollination method

Flower buds of L. X ‘formolongi used as ovary parents were

emasculated one day before anthesis and the stigma was cov-

ered with aluminum foil. The cut-style pollination method(12)

was applied as follows. When stigmatic exudates appeared，

the style was cut off 1， 3， or 5 cm above the ovary. Then a

5-mm longitudinal incision was made in the remaining part of

the style. After mounting the desired pollen into the slit， the

styles were covered again with aluminum foil.

Ovary-ovule culture

Ten days after pollination， ovaries were harvested and sliced

into 12 sections after surface sterilizatIon. Positions (top to

base) of each section on the ovary we陀 recorded.Then the sec-

tions were placed on MS medium supplemented with 1.0 mg 1-1

NAA， 100 g 1-1 sucrose and 7 g 1-1 agar (medium 1) (玖 After

40 days of incubation on medium 1， all ovules except browned

ones were transferred to MS m巴diurnsupplemented with 0.1

mg 1-1 NAA， 50 g 1-1 sucrose and 7 g 1-1 agar (medium n).

All cultures were kept at 250

C in dark conditions. The germi-

nation frequency was evaluated after 150 days of culture on

medium 11.

Acclimatization and growing conditions

Germinated ovules were transferr吋 toa medium containing

3 g 1-1 Hyponex (complete soluble fertilizer， N: P2U5: K2Uニ 16:

10: 5; Hyponex Co. Ltd.， Japan) and 7 g r1 agar (H medium)

and cultured under light conditions at 20μmol m -2 S -1 PPFD

with a 16 hr photoperiod for another several weeks. The ob-

tained plantlets with 2-3 leaves were acclimatized and grown

in a greenhouse at a minimum temperature of 12 OC until first

flowering. The developed bulbs were then grown in an unheat司

ed gree凶louse.

Analysis of obtained progenies

The ploidy level of putative hybrids was determined using

flow cytometry (FCM) (Ploidy Analyzer PA， Part巴c1nc.，

Germany) . About a 5 mm length of young leaf tissue was col-

lected from the plantlet. Sample preparation and measurement

conditions were identical to those described by Fukai et al. (14) •

The diploid L. X formolongi was used as an intemal standard.

To determine the pollen fertility ofthe putative hybrids， pol-

len was placed on a medium containing 40 g 1-1 sucrose and 5

g 1-1 agar. Then pollen was incubated at 25

0

C ovemight.

The pollen surface structure was observed using a scanning

electron microscope (SEM， S-2150; Hitachi Ltd.). Dried pol-

len was mounted on an observation stage and coated with Pt

before SEM observations.

Restoration of pol恰ngermination using high-tempera-

ture treatment of flower buds

Plants of six independent LT hybrids obtained in this study

were grown in an unheated greenhouse. Wh巳nflower buds be-

came visible， the plants with variously sized flower buds were

transferred to a growth chamber (with solar irradiation) that

was controlled for a week at 300

C ， as described in Natenapit

et al. (j2). A total of 57 flower buds from 18 plants of six lines

were subjected to high-temperature tr巳atment.Thereafter， the

plants were taken back to the greenhouse， where they were

kept until flowering. Pollen from each flower was harvested

and it was then subjected to poll巴ngermination tests by use of

a medium contairiing 60 g 1-1 sucrose and 5 g 1-

1 agar. Pol-

len showing germination ability was used in further crossing

with L. X戸rmolongiusing the cut-style pollination method

described above.

Chromosome preparation and genomic in situ hybrid-

ization (GISH)

Root tips were collected and kept in distilled water in a tube

on ice for 24 h and fixed in ethanol acetic acid solution (3 : 1)

for 12 h. The root tips were incubated in a pectolytic enzyme

mixture containing 1% (w/v) pectolyase Y-23， 4% (w/v) cel-

lulase RS， 75 mmol KCI， and 7.5 mmol Na2 EDTA (pH 4)

S. Fukai et al. : Production of Interspecific L-T Hybrid Lilies 13

at 370

C for 1-2 h. Squash preparations were made in a drop

of 45% acetic acid and frozen at _80oC . The cover slips were

removed using a razor b1ade. Slides were dehydrated in an

ethano1 series (70%-85%-99% vo1.) at 5 min interva1s and

air-dried.

Genomic DNA of L. X formolongi (‘White 1ancer') was

used as probe， 1abe1ed with b則 in-14-dATP(BioNick™

Labe1i時 System;Invitrogen Corp.) usi時 nicktrans1ation ac-

cording to the manufacturer' s instructions. The hybridization

mixture consisted of 50% (v/v) deionized formamide， 10%

(w/v) dextran su1fate， 2 X SSCラ and25 ngμ1-1 biotiny1ated

genomic DNA. To each slide， 10μ1 ofthe hybridization mix-

ture was app1ied， covered with a cover slip， and sea1ed with

rubber cement. The slides wer巴 denaturedon a hot p1ate at

78t for 30-45 s and incubated at 37t overnight (16-18 h).

After peeling off the rubber cement， the covers1ips were

removed in 2 X SSC. The slides were then washed in two

changes of 50% (v/v) formamid巴 at37 oC for 10 min each，

and twice in 2 X SSC at room tempera旬re.Following a short

rinse with 4 X SSC and 0.05% (v/v) Tween 20， and b1ock-

ing by drop of 3% (w/v) BSA for 5 min at room temperature，

30μ1 of 3% BSA solution and streptavidin Cy3 was app1ied

to each slide. A cove白r司叫i切pwa出sapplied and t白h巴 slides w 巳訂re

incubated for 1 h a幻t3rc. The slides were then rinsed four times with 4 X SSC and 0.05% (v/v) Tween 20 at room tem-

perature for 5 min each. After a short rinse with 2 X SSC，

the slides weぽr巴nns

DAPI and an凶凶tifiぬ"ad巴 (DABCO) were appli 巴d to it. Before ob-

servation using a microscop巴 (AX70FL; Olympus Corp.)，

the slides were incubated at room temperature for at 1east 1 h.

Results and Discussion

Genotype differences in progeny production

The 1ength of the remaining sty1e in cut“sty1e pollination

method affected the germination frequencies of cultured

ovu1es in three LT crosses (Tab1e 1). A 10nger sty1e (5 cm)

produced a higher number of germinated ovu1es than those

of shorter sty1e (1 cm). The pollen tube growth is suspended

in the sty1e in interspecific crossing of Lilium. Therefore， the

shortened distance from the pollen germination site to 0刊 1es

(cut-sty1e pollination) pr吋 ucesferti1ization (1). However， a

shorter sty1e 1ength did not a1ways produce better resu1ts in

LO crosses U3)， equa1 to those described herein. In further LT

crosses in this experiment， a 5 cm sty1e 1ength was used.

Germination frequencies varied depending on pollen par-

ents and the crossing year (Tab1e 2). All tested pollen parents，

including Aurelian hybrids and wi1d species， produced prog-

巴nies.The highest frequency of ovu1e germination was 1.1%

in the crossing of L. X formolongi‘White Lancer' with‘Pink

Trumpet' in 2006. Some cross combinations were done for

three years. Their productivities differed considerab1y by year.

Most cross combinations showed 0-0.2% of ovu1e germina-

tion. The frequency was equiva1ent to the crossings of L. X

戸rmolongiwith Asiatic trumpet white 1i1y species (7).

Germinated ovu1es appeared at various positions on the

ovary (Tab1e 3)， indicating that the pollen tube reached all

parts of the ovary. This result suggests that the 1imitation of

fertilization was not simp1y the distance from pollen to ovu1e.

Even though the pollen tube reached the ovary by cut sty1e

pollination， pollen tubes often ignored micropy1es when the

sty1e 1ength was short in compatib1e crosses (15). Recently， the

substance acting as the directiona1 cue for pollen tube growth

has been identified (16). However， the manner in which the

Tab1e 1 Effect of sty1e 1ength on number of germinated ovu1es in LT crossings *.

Pollen parents** length of style No. cultured ovaries No. cultured ovules

5cm 8 4101

Pink seedling 3cm 7 3622

1cm 6 2563

5cm 6 2982

Apricot seedling 3cm 8 4477

1cm 7 3232

5cm 9 5197

A仕icanQueen 3cm 8 4335

1cm 6 2681

* Ovary parent is‘Augusta'. * * Pink seedling: se巳dlingof ‘Pink perfection'， Apricot seedling: seedling of apricot flowered breeding line.

*キ*Only cullus r巴gen巴rated.

No. germinated ovules (%)

5 (0.12) 2 (0.06) o (0.00) 7 (0.24) 1 (0.02) o (0.00) 2 (0.04) 2 (0.04)村*

o (0.00)

14 Tech. Bul1. Fac. Agr. Kagawa Univ.， Vo1. 65， 2013

Table 2 Genotype differences in production ofhybrids in LT crossings

Pollen parents Ovary parents * No. cultured ovaries No. cultured ovules No.g巴rminatedovules (%) White Henry (07) * * 明/ 8 *** 4 (-) White Henry (08) w 8 4499 o (0) White Henry (09) 、J 5 2740 25 (0.9)

Pink Perf(巴ction(07) W 7 13 (一)Pink Perfection (08) W 7 3403 8 (0，2) Golden Sprend巴r(07) W 5 o (0) Golden Sprender (08) W 16 10713 15 (0.1) Golden Spr巴nder(09) W 6 3442 22 (0.6) Apricot seedling (08) W 10 5831 4 (O.l) Green Palace (07) W 9 22 (一)Green Palace (07) A 2 1235 3 (0.2) Green Palace (09) W 10 5163 15 (0.3) A仕icanQue巳n(07) A 2 1159 1 (02) African Queen (09) W 6 2908 9 (0.2)

L. henry (08) 、J 10 6309 19 (0.3) L. henry (09) 、J 6 3411 17 (0.5)

L ，centifolium (y) (08) W 10 5344 8 (0.2) L. centifolium (y) (09) W 6 3298 5 (0.2)

Pink Trumpet (06) W 10 2310 26 (1.1) Pink Trumpet (07) A 1 506 o (0)

*Ovary parent was L. X formolongi， A・'Augusta'，W;‘White Lancer'. **r しrossmgyear

料*Data was lost accientaly.

Table 3

Positions in the ovary *

Top 1

2

3 4

5

6

7

8

9 10

11

Base 12

Number of germinat怠dovules in each position of ovary in some LT cross combinatuions，

Pollen parents (Iength of style) * *

Pink (5) Pink (3) PP (5) GP (5) LH (5) WH (5)

•••• ••• • •• •• • ••• ••• • • • • • •••• • • • • • • •• •• • •• • •••••• • ••• • • • • •••• • • ••••• ••• • ••••• Total number of cultured ovules 4101 3622 3403 5163 3412 2744

キ'Ovarywas devided to 12 cross sections and positions in an ovary was recorded，

* *Pink: Pink seedking， PP: Pink Perfection， GP: Green Palace， LH: L. henry， WH: White Henry. • represents a germinated ovule.

Characteristics of obtained L T hybrids

Som巳progeni必esflowered du凶lrlll

interaction of the pollen tube and ovule play a role in interspe-

cific crossing with shortened distance from the pollen to ovule

remains unclear.

Germinated ovules developed a cotyledon and roots noト

mally. Some ovules produced calli or abnor百lalswollen leaves

followed by shoot formation on medium II. The shoots devel-

oped plantlets after transferring t0 H medium; they acclima-

tized successfully

moωs託叫tlyi凶nthe second spring from a叫cclima瓜凶t“i包za瓜叩t“iぬonin the heated

gr巳∞巴nhouse(minimum t印emp巴rat旬ur閃eof 120

C )， These hybrid出s

flowered during early May to earぢJunein the heated green-

house and during late May to mid-June in the non-heated

greenhouse. The short juvenile phase (flowering within a year

from crossing) of some LT hybrids is derived合omL. X for-

molongi， which can flower in 8 months from seeds. The short

juvenile phase of interspecific hybrids has also appeared when

S. Fukai et al. : Production of Interspecific L-T Hybrid Lilies 15

Fig. 1 Flowers ofLT hybrids. Progenies of L. x戸rmolongiand ‘Pink se巳dling'A:LP5-4，B: LP5-3，C: LP5-8

Fig.2 Micrograph ofpollen surfaces in interspecific hybrid and parents

A: L. X folmolongi; B: Aurelian hybrid‘Pink Perfection' seedling;

C: hybrid of A and B. Bars indicate 10μm

L. formosanum， a parent of L. X formolongi， was used as an

ovarγparent(17). The lack of a great difference in flowering

time of LT hybrids between heated and non-heated green-

house suggests a rather low chilling requirement for flowering

of the LT hybrids. In this study， L. X formolongi‘August' was

mainly used as ovule pa民 nts.Reportedly，‘Augusta' has a lower

chilling requir・'eme凶 forflowering among the L. X戸rmolongi

genotypes (18) •

The morphology of flowers was intermediate of parents，

with considerable variations in perianth width and length (Fig.

1). Prog巴niesderived from crosses of different flower shape

genotypes often show a disordered flower shape， i.e. cleaved

flower tubes. Some LT progenies also had disordered flowers.

The perianth color was also intermediate of parents， resulting

in a pale color. Progenies of L. X formolongi and ‘Pink Per司

fection' seedlings showed that the perianth exterior was pale

pink to pale pinkish-brown; the interior was pale green with a

yellow throat (base ofthe corolla tube)

Mother-resembling plants were obtained through crossing L.

X formolongi with ‘Golden Splendor'. The mother-resembling

plants had white trumpet-shaped flowers with fertile pollen，

suggesting that they are apomicts. Production of apomicts was

reported when L. X formolongi was used as an ovary parent

in interspecific hybridization (13， 191.

FCM analysis showed that all obtained progenies were

diploids (data not shown). Pollen fertility of the LT hybrids

was mostly zero. However， pollen with low germination abiト

ity (0.1-1.9%) was observed sporadically. Results of SEM

observations revealed that most pollen of the LT hybrids had

shrunk， but round pollen was also observed at a very low fre-

quency. The plants in the genus Lilium had pollen with a re-

ticulated surface. L. X formolongi had large reticulation with

a smooth line， whe児巴r陀巳aωsT hybrid had rather small r閃巴d凶tlCU叫!la瓜tiωon1

with a bead-like shape (Fi氾g.2)ト.The obtω削a幻inedLT hybrid had

inte白n宅'm

During 4 y巴arscultur印巳 af武ie白racclima抗ti位za瓜11ωonラ numbers of

Tech. Bull. Fac. Agr. Kagawa Univ.， Vol. 65， 2013 16

Crossing of L. xルrmo!ongiand LT hybrid by use of fertility restored pollen

Ovary parents No. acclimatized plants

90一0

0一0

0

No. genninated ovules (%)

13 (0.9)

2 (0.1) 。。0

0

No. cultured ovules

1480

2049

2382

1227

1278

1686

Pollen p31官nts(pollen genn川 ation%)

LP5-4-1-1 W 3 (8~ A 4

LP5-4-1-2 W 4 (3.6%) A 3

LP5-4-2-2 W 2 (3.5%) A 4

Ovary parents were L. X formolongi. A:・Augusta'，W・'WhiteLancer'

No. cultured ovaries

Table 4

bilities: the us巴dLT pollen that restored gerl11ination ability

by high tel11perature was an unr巴ducedgamete (2川，or the

ovary parent (L. X .forl11o!ongi) produced an unreduced egg

and the LT pollen was 11 level. The genomic constitution of

four independent BC1 was analyzed using GISH. All tested

BC1 had 36 chromosomes: three of them had 12 T and 24 L

chromosomes; one (LT 351) had 13 T and 23 L chromosol11es

(Fig. 4). Results show that the genomic const山 tionof the

triploid BC1s was LLT. Furthermore， the BC1 progenies had

3-7 chromosomes with homoeologous recombination (Table

5). Each recombinant po引tionwas different in a plant. Th巳

results indicate that the pollen with restored fl巴rtilityby high-

temperature treatment was an unreduced gamete， and that it

was produced by both the first division restitution (FDR)

and indeterminate meiotic restitution， as described by Karlov

el al.1却

)and Lim et al. 121 l• Lack of introgression caused by

absence of recombination is a severe limitation for creation

bultヲinLT hybrids (LP5-3， LP5-4， LP5-8， LP5← 12) in-

cr巴ased9， 8， 25， 10， respectiv巴ly.The result indicates that the

propagation rates by natural division of LT bulbs were 2 to 2.2.

Restoration of pollen fertility and back cross

After high-temperature treatment， 12 of 57 nower buds

derived from five LT progenies showed restoration of pollen

germination capability. The pollen germination仕equency

was 0.1-8% depending on the now巴rbuds. The germination

frequency remain巴dconsiderably lower than that of either

parental plant. Pollen from three indep巴ndentplants showed

rather high pollen germination ability: LP5-4-1-1 (8%)，

LP5-4-1-2 (3，6%)， and LP5-4-2-2 (3.5%) were sel巴cted

and used for backcrossing. The pollen was mounted on cut-

styles of L. X .forl11olongi・Augusta'and 'White Lanc紅 白Only

the pollen showing the highest germination (LP5-4-1-1)

produced BC1 progeni巳s.Ovule germination was 0.9% when

'Wh山 Lancer'was the ovary par巴nt(Table 4); it was equiva- of variations. Use of an unreduced gamete is the only way to

lent to LT crosses.

Characteristics of obtained LLT hybrids (BC1)

Results of FCM analysis showed that all tested BCl prog-

el1les w巴retriploid (Fig. 3). The reslllts present two poss卜

150 Count

100

50

Chromosome constitlltion analyzed using GISH in LLT progenies. DNAoぱfL. X j戸ρbかrm〆-mη01わon噌glパ(‘ Whi巾t陀el川a創加nc叩E印rつwaωs use吋d as prob 巴 labelled wit山hbiotin (p戸3別刊川i日川nkn仏nlloωr巴scenc 巴)入， unlabelled Trllmηlpet hybrid w司ascount巴rstainedwith DAPI (blue fluorescence). Arrows indicate recombi-natIon sltes.

Fig.4 。o 50 100 Relative tlruorescence intensity

FCM analysis of progeny A: diploid L. X .forl11010ngi B: progeny of L. X .forl11olongi X LT

Fig.3

s. Fukai et al. : Production ofInterspecific L-T Hybrid Lilies

Table 5 Number of chromosomes and chromosome consti-tution of LLT hybrids

Progeny Chromosome constitution

No. r官combinantchromosomes

(No. recombination sites) L T

1 (1) 2 (2)

2 (2) 3 (3)

4 (4) 2 (2)

3 (5) 4 (4)

LT351 LT371 LT391 LT481

L T

23 13 24 12 24 12 24 12

，d

vi

hu vd

LH

TI

YL

YL

fI

o

fi

pLV W

G

I

E--A E1u

ob

.-ui

resolve this problem. Results show that the unreduced gamete

was induced by high-temperature treatment in LT hybrids.

High-temperature treatment is a simple method using no spe-

cific equipment(22J and no chemicals叫 toinduce an unreduced

gamete.

Since obtained hybrids had two sets of L genome they

produced perfect trumpet-shaped flowers (Fig. 5)， as demon-

strated in LLO hybrid (24).

Results show successful crosses of L. x戸rmolongiand

Trumpet hybrid lilies and the overcoming of sterility through

production ofunreduced gamete using high-temperature treat-

ment， ther巴byensuring the production of this n巴wtype ofhy-

brid lily.

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S. Fukai et al. : Production of Interspecific L-T Hybrid Lilies 19

新テッポウユリとトランペットハイブリッドの遠縁交雑ユリの作出

深井誠一・ジットラム ネイナピット・望月麻美・花園E由美・鳴海貴子

要 約

花柱切断法および、子房目玉珠培養を用いて，新テッポウユリ (L. X戸rmolongi) (L) を子房親としてトランベットハ

イブリッド (T) との遠縁交雑種の作出を行った試されたすべての交配組合せで後代が獲得できたが，培養した伍珠

の発芽率は花粉親であるTの遺伝子型と交配年により異なった.得られたLT後代は，両親の中間型の形態を示し，低

い花粉稔性を有していた菅の高温処理により交雑種の花粉稔性を回復させ，それを新テッポウユリに戻し交配したと

ころ，三倍体の後代が得られた.ゲノミックインサイチューハイブリダイゼーションにより，得られた後代のゲノム構

成はLLTであり，いくつかのLTゲノム聞の同祖組換えを有していることが明らかとなった.このことから高温処理に

より稔性回復したLTの花粉は，非還元性花粉であることが示された