全 文 ：　遗 传 学 报　Ac ta Gene tica S in ica, November 2005, 32 (11):1191～ 1198 ISSN 0379 -4172
收稿日期:2005 - 04 -07;修回日期:2005 -05 -28
基金项目:国家自然科学基金项目 (编号:30370143, 30270651)[ Supported by the Nat ional Natural Sc ience Foundat ion o f China (No.
30370143, 30270651)]
作者简介:宿红艳(1976 -),女,博士 ,研究方向:植物发育生物学
①　通讯作者。 E-ma il:zhangxs@ sdau. edu. cn
Characterization and Expression Analysis of a MADS Box Gene,
HoMADS 2, inHyacinthus orientalis L.
SU Hong-Yan1, 2 , LIQuan-Z i1 , LI X ing-Guo1 , ZHANG X ian-Sheng1, ①
(1. Shandong Key Laboratory ofC rop B io logy, Co llege o fL ife Sc iences , Shandong Agricultura l
Univers ity , Taian　271018, China;
2. Co llege o fL ife Sc iences , Yantai Norma lUniversity, Yan ta i　264025, China)
Abstract:A MADS box geneHoMADS 2 w as cloned fromHyac in thus o rien ta lis L. in th is study. Sequence com-
pa rison revea led tha tHoMADS 2 was h igh ly hom o logous to the class B MADS box genes. Fu rthe rmore, phy loge-
ne tic ana lysis show ed tha tHoMADS 2 was close ly re la ted toP I gene fam ily and th is w as a lso suppo rted by the
p resence o f spec ific d iagnostic sites o f P I hom o logs in K box dom a in and C te rm ina l reg ion, sugges ting tha tHo-
MADS 2 m igh t be aP I-like gene. HoMADS 2 mRNA was accumu la ted in a ll flora l organs, d iffe rent from the ex-
p ress ion pa tte rns o fPI homo logs in d ico ts. D ur ing in vitro flower deve lopmen t,HoMADS 2 expression was con-
s titu tive ly exp ressed and no t affected by the p resence o f cytok in in and auxin in the regenera ted flowers. Our re-
su lts ind ica ted that the express ion ofHoMADS 2 is d iffe rent from those o f bo thHAG 1 andHoMADS 1 in respon-
d ing to p lan t ho rmones du ring in vitro flow er deve lopmen .t
K ey words:HoMADS2;ho rmone;Hyacin thus o rien ta lis L.;in v itro flow er
风信子 HoMADS 2基因的克隆及表达分析
宿红艳 1, 2 , 李全梓 1 , 李兴国 1 , 张宪省 1, ①
(1. 山东农业大学生命科学学院 , 山东省作物生物学重点实验室 , 泰安　271018;
2. 烟台师范学院生命科学学院 , 烟台　264025)
摘　要:利用同源克隆策略 , 从风信子中分离出一个 MADS box基因 ,命名为 HoMADS 2。序列比较分析表明 ,
HoMADS2与 B类 MADS box蛋白具有较高的同源性。分子进化树分析显示 , HoMADS2与 P I家族类聚在一起。
同时 , 在 HoMADS2的 K box和 C末端区域均具有 P I家族的特征序列。以上序列分析结果表明 ,HoMADS 2可能
是 P I的一个同源基因。 RNA分子杂交结果显示 ,H oMADS 2在四轮花器官中均表达 ,其表达模式不同于双子叶植
物中 P I同源基因。利用风信子离体花器官再生系统研究表明 , HoMADS 2在再生花芽中的表达不同于 HoMADS1
和 HAG 1,该基因在再生花芽发育过程中组成型表达 , 不受外源细胞分裂素和生长素的影响。
关键词:HoMADS2;激素;风信子;离体花发育
中图分类号:Q943. 2　　　文献标识码:A　　　文章编号:0379-4172(2005)11-1191-08
　　Analysis of homeotic mutants inA rabidops is
and Antirrh inum led to the proposa l of an ABC
model to exp la in how the fate of a f lo ra l organ pri-
mord ia is determ ined. Accord ing to th ismodel, the
identities of flora l organs are spec if ied by the func-
tions of three c lass genes A, B and C. Each class
funct ions in two adjacentwhorls of the flower
[ 1, 2]
.
O f them , class B genes, such as AP 3
(APETALA 3) andPI (P IST ILLATA) in A rab idop-
s is, play key ro les in spec ify ing peta l and stamen
identities
[ 3 , 4]
.
AP in in in in in in in in genes encode prote ins
conta in ing an N-term ina lDNA bind ing MADS box
domain , an I domain , a K box domain and a d iver-
gentC term inus
[ 5 -8]
. D ifferent from the c lasses A
and C genes in form ing hom odmi ers , AP 3 and P I
were thought to func tion together as heterodmi -
ers
[ 3, 4, 7]
. So far ,many ortho logs of AP 3 and P I
have been identified and charac terized and they
appeared to func tion in a smi ila rmanner in d iffer-
ent p lant spec ies
[ 9 -11]
.
Based on phy logenetic ana lysis , genes of
AP 3and P I fam ilieswere thought to arise from an
ances tra l gene through duplica tion , and the AP 3
gene lineage underwentanotherdup lica tion ,which
gave rise to two AP 3 sublineages:euAP3 and
TM6
[ 8, 12, 13]
. However ,whether there is another d i-
vergence in the sequences of P I lineages during
evo lu tion is still unclear. Each of these lineages is
characterized by a series of specific d iagnos tic
s ites in the MADS box ,K box and C domains , al-
though they share overa ll sequence smi ila rity w ith
each other. For examp le, PI lineage possesses a
short hydrophob ic reg ion named PI motif a t the C
term inus ,wh ich is a lso present in the C-te rm ina l
reg ion of the prote in of the B func tiona l ancestra l
gene.W hereas in the AP 3 lineage , the P I-motif
underwent sequence divers ifica tion and gave rise
to P I-derivedm otif[ 8, 14] . I twas shown that the line-
age-spec ific motifs in the C term inus are critica l
for the func tiona l d ivers ifica tion of these gene line-
ages ,and the changes in the func tions of the en-
coded produc ts may contribute to the d ivers ity o f
flora lmorphology
[ 15]
.
F lowers ofmonocots are d iffe rent from those
ofd icots inmany aspec ts , such as in themorphol-
ogy of sepals and peta ls. Hyac in thus (Hyac inthus
orienta lis L. ),a monocot has two whorls of tepa ls
resembling peta ls instead of sepa ls and peta ls in
d icots. A hyac in thus in v itro flo ra l regeneration
system was desc ribed in a previous paper , in
which the types of regenerated flora l organs were
exac tly contro lled by the leve ls of p lant hor-
mones
[ 16, 17]
. A t h igh leve ls of cy tok in in and aux in
in the m edia , only tepa l was d ifferentia ted in re-
generated flowers.W hen the flowers transferred to
the med ia w ith low leve ls of cy tokin in and auxin ,
stamens and carpe ls w ith ovu les were produced
from regenera ted flowers. Th is system is very
usefu l to s tudy the molecu larmechanism of flo ra l
organ development regu la ted by p lant hormones.
O ur prev ious stud ies showed tha t the c lass C
gene HAG1 (Hyac inthus AGAMOUS1) and the
class D gene HoMADS1 (Hyacin thus orienta lis
MADS box 1),which are homologs of AG (AGA-
MOUS)and AGL11 (AGAMOUS-Like 11) in Ara-
b idopsis , respec tively , were induced by plant hor-
mones during in v itro flower deve lopm ent
[ 18 -24]
.
HAG1was induced by high leve ls of cy tokin in and
aux in during in itia tion of hyac in thus flowers in
v itro , whereas HoMADS1 was induced by low
leve ls of cytok in in and aux in which were responsi-
b le for ovu le forma tion
[ 21, 24]
. I t is like ly that the in-
duction of the homeotic genes by hormones is re-
quired for the d ifferentia tion of flora l organs in
v itro. In th is paper, we iso la ted a hyacin thus
MADS box genewhich shares h igh homology w ith
PI in Arabidops is and its expression patterns in
p lanta and in regenerated flowers were ana lysed.
1　Ma teria ls and Methods
1. 1　Plantmateria ls
Bulbs ofHyac in thus orienta lis L. (cv. Delft
b lue) were purchased from J ind i Company in
1192 遗传学报　Acta Gene tica S in ica　Vo.l 32　No. 11　2005
China. The bulbswere p lanted in the fie ld at Uni-
vers ity campus in October for 3 months, and then
were used as expermi enta lmater ia ls.
1. 2　Preparation o f the exp lants and floral
bud regeneration
　　The bulbs were d issected, and the tepa ls of
flora l buds at the stage of b ice llu lar po llen were
se lec ted as exp lants for flo ra l bud regeneration.
The preparat ion of cu lture med ia and procedures
of f lo ra l bud regeneration were performed as de-
scribed prev iously
[ 16, 17]
.
1. 3　 Iso lation o fHoMADS 2
Tota lRNA was iso lated from flora l tissues by
using the to ta l RNA iso la tion system (P romega
Corporation, USA ). The RNA was used for re-
verse-transcrip tase polymerase chain reaction
(RT-PCR) and Northern hybrid izat ion analysis.
To iso la te the P I homolog from hyacin thus, tota l
RNA of young tepa ls was used as templates for
RT-PCR by a degenerated prmi er 5′-GG(A /G)
AG(G /A)GG(G /A)AA(A /G)AT(T /C)GA(G /
A)AT(T /C /A)AA-3′and a prmi er o ligo(dT). RT-
PCR was performed as described prev iously
[ 21]
.
Comparison w ith sequences in the databases re-
vea led that one fragment showed h igh homolog
w ith class B MADS box genes. To iso la te the
complete 5′reg ion of th is gene, 5′RACE PCR
was carried outw ith the gene-spec ific prmi er(5′-
GAGCGCCAGTTACGCCATTC-3′) and an a-
bridged universa l amplifica tion prmi er accord ing to
manufacturer’ s instructions (G ibco BRL K it,
UK). PCR was carried out as fo llows:94℃ fo r 3
m in, fo llowed by 35 cyc les of 94℃ fo r 1 m in, 56℃
for 1 m in and 72℃ fo r 2 m in.
To get the fu ll-length cDNA sequence of th is
gene, RT-PCR was carried out using 5′and 3′
spec ific prmi ers, respec tive ly. The PCR thermal
cyc les were carried out as fo llow ing:94℃ fo r 3
m in, fo llowed by 35 cyc les of 94 ℃ fo r 1 m in,
60℃ for 1 m in and 72℃ for 2 m in.
PCR fragments were cloned in to pGEM-T
vectors (Promega Corporat ion, USA) and se-
quenced w ith an ABI PR ISM
TM
377 DNA Sequen-
cer (Perk in E lemer, USA). The sequence data
have been subm itted to the GenBank databases
under access ion number AF134114 and the gene
was designated asHoMADS 2.
1. 4　Northern hybridization
Twenty m icrograms of to ta l RNA were frac-
t ionated by e lec trophoresis in 1. 2 % agarose gel
conta in ing forma ldehyde and were transferred
from agarose gels to nylon memebranes as de-
scr ibed by Sambrook et a l.
[ 25]
. P rehybrid izat ion
was performed at 42℃ fo r 12 h. The probe
spec if ic for HoMADS 2 was the 3′reg ion of its
cDNA which was amplified by two spec ific prmi -
ers:SP1 5′-TGGCATATTTACTGCATCATCAG-3′
and SP2 5′-ATCTAATGACGATAATCATTCAG-
3′. Hybrid izat ion was conducted at 42℃ fo r 48 h.
F ilte rs were washed subsequently in 2 × SSC
w ith 0.2% SDS and 0. 2 ×SSC w ith 0. 2% SDS.
Autorad iographywas performed at -70℃.
2　Results
2. 1　Morpho logy o f hyacinthus flowers in
p lanta and in vitro
　　Hyacin thus flower cons ists of four whorls of
flo ra l organs. The outer two whorls have three te-
pals. The th ird whorl and fourth whorl have six sta-
mens and a gynoec ium , respective ly (F ig. 1, A).
Continuous d ifferentia tion of tepa ls was induced
from regenerated flora l buds in themed ium conta i-
n ing h igh leve ls of cy tok in in and aux in (F ig. 1,
B). However, the stamens could be induced when
the regenerated flowers were transferred to the
med ia w ith low leve ls of cytok in in and aux in (data
not shown ). Deta iled descrip tions of in vitro
flowers can be found in our prev ious stud ies
[ 21]
.
1193SU Hong-Yane t a l.:Characte r iza tion and Exp ression Ana lys is o f a MADS BoxGene…
F ig. 1　Flowers of hyacinthus in
p lanta and in v itro
A:Long itudinal sect ion of a flower in p lanta, pa rt flora l organs
be ing removed;B:A regenerated flow erw ith a few whorls of
tepals at high level hormones. gy:gynoec ium;st:stamen;te:
tepa.l Bar =1 mm.
2. 2　 Iso lation o fHoMADS 2 from hyacinthus
A few MADS box genes were iso la ted from
the flora l tissues of hyac inthus. Sequence compar-
ison revealed that one of them shared high homol-
ogy to the c lassBMADS box genes, such asP IS-
T ILLATA in A rab idopsis and was des ignated as
HoMADS 2 ( fo rHyac in thus orienta lis MADS box
2).HoMADS 2 cDNAwas 888 bp long encod ing a
202 am ino acid prote in. A lignment of am ino acid
sequence ofHoMADS2 and the representa tives of
puta t ive B group prote ins revea led severa l fea-
tu res (F ig. 2). F irstly, HoMADS2 shared higher
F ig. 2　A lignmen t of HoMADS2 w ith the re la ted MADS box prote ins
HoMADS2 is aligned to putat ive proteins:OsMADS2 and OsMADS4 o f rice (Chung et a l. 1995), OsMADS16 (AAD19872) of
rice,WP I(BAD12462) ofw heat, P I(P48007) and AP3 (P35632) ofA rabidops is, GLO (Q03378) and DEF (P23706) of An-
t irrh inum. The f irs t underlined reg ion isMADS box doma in and the second underlined reg ion is K box dom ain. The firs tboxed re-
g ion (KHENL) represents a highly conserved sequence (KHExL) among P I gene fam ily and the second boxed reg ion (MAL-
RVQPVQPNLQE) contains consensus sequences for P I-mot if (MPFxFRVQPxQPNLQE). Conserved Glu-97 and G ly-131 are
marked w ith an as terisk. Dark shading w ith wh ite letters and g ray shading w ith dark letters re flect 100 and 50% sequence con-
serva tion, respect ively. Gaps introduced to mi prove alignm ent are indicated by dashes.
1194 遗传学报　Acta Gene tica S in ica　Vo.l 32　No. 11　2005
F ig. 3　Phy logenetic tree of the MADS box genes
This phy logenet ic analys is shows thatHoMADS2 is c losely re lated to three monocotsP Ihomo logsW PI(wheat), OsMADS2 and
OsMADS4 ( rice). Nam e of the hyac inthus MADS box prote in HoMADS2 is underlined. The tree groups inc lude:WPI
(BAD12462) of w heat;OsMADS2 and OsMADS4 of rice (Chung et a l. 1995);OsMADS16 (AAD19872) of rice;HAG1
(AF099937), HoMADS1 (AAF088030) and HoMADS2 (AF134114) o f hyac inthus;APETALA1 (P35631), SEP1 (P29382),
SEP2 (P29384), SEP3 (O22456), P I(P48007), AP3 (P35632), AGAMOUS (P17839), SHP1 (P29381), SHP2 (P29385),
and AGL11 (Q38836) of Arab idops is;FBP1 (Q03488), FBP3 (S60288) and PhTM6 (AAS46017) of petunia;NTGLO
(Q03416) of tobacco;AmFUL (AAP83363), GLO (Q03378) and DEF (P23706) of Antirrhnium;SLM3 (CCA56657) of
S ilene lat ifo lia;GDEF1 (CCA08802) and GDEF2 (CCA08803) ofGerbera hybrida;LMADS1 (AAM 27456) and LMADS2
(AAS01766) of lily. The rooted tree is genera ted w ith the d is tance matrix us ing the Neighbor-Joining method (Sa itou and Nei,
1987) of the DNAMANMASED prog ram. The length of the branch line indica tes the extent ofd ifference accord ing to the sca le at
upper left and num bers on major branches ind icate bootstrap estmi a tes for 1 000 replicate ana lys is.
homologyw ith PI homologs, such asWP I in wheat
(62% identity), OsMADS2 (60% ident ity) and
OsMADS4 in r ice (59% identity), P I inA rabidop-
s is (53% identity), and GLOBOSA in Antirrh inum
(54% ident ity) than AP3 homologs inc lud ing O s-
MADS16 (39% identity) in r ice, AP3 in A rab idop-
s is (36% ident ity) and DEF IC IENS inAntirrh inum
(39% identity)[ 2, 4, 10, 26 -28] . Secondly, w ith in K box
domain, a sequence (KHENL)matched perfectly
to a h igh ly conserved sequence (KHExL) in P I
homologs. The comparab le sequence in the same
reg ion of the AP3 homo logs is Q /HYExM
[ 8]
. Be-
sides, the positions 97 and 131 were G lu and G ly
w ith in K box domain of HoMADS2, respect ive ly,
and both of them were unique am ino ac ids in P I
homologs. W hereas in AP3 homologs, the spec ific
residueswere Arg-102 and Ser or Thr-131[ 11, 29] .
F ina lly, w ith in the C term ina l reg ion ofHoMADS2,
1195SU Hong-Yane t a l.:Characte r iza tion and Exp ression Ana lys is o f a MADS BoxGene…
the sequence MALRVQPVQPNLQE showed 68%
identity to P I mot ifMPFxFRVQPxQPNLQE wh ich
was conserved in P I fam ily, and also showed 54%
identity to P I-der ived mot if FxFRLOPSQPNLH
which was found in AP3 gene fam ily
[ 8]
. In add i-
tion, a phy logenetic tree of the re la ted MADS box
genes was construc ted based on the am ino ac id
sequence alignment of P I and AP3 homologs
(F ig. 3). The dendrogram shows that the HoM-
ADS2 prote in is close ly re la ted to O sMADS2 (a
P I homolog in rice), OsMADS4 and WP I in P I
gene group
[ 26, 28]
. Thus, these resu lts suggested
thatHoMADS 2 m ight be aPI-like gene.
2. 3　HoMADS 2 was exp ressed in all flo ral or-
gans
　　To explore the tempora l and spat ia l expres-
s ion pattern ofHoMADS 2 in the vegetat ive and
flora l organs in hyac in thus, the analysis of RNA
blot hybrid izat ion was performed. As shown in
F ig. 4,HoMADS 2 transcrip tswere not detected in
vegetat ive organs inc lud ing roots, sca le leaves
and leaves. In contrast, HoMADS 2 mRNA was
accumulated in a ll flora l organs includ ing tepa ls,
stamens and carpe ls. The leve ls of HoMADS 2
mRNA in carpe lswas predom inantly h igh unt il the
completion of flower deve lopmen.t Th is pattern is
d iffe rent from that o fP I homologs in d icots, wh ich
is prmi arily expressed in the second-and th ird-
whorl flo ra l organs
[ 4, 10, 30]
.
2. 4　HoMADS 2 is not induced by cytokin in
and auxin in regenerated flowers
　　To study whetherHoMADS 2 expression is
induced by cytokin in and aux in dur ing the deve l-
opment of regenerated flowers, we tried to use the
in v itro f lo ra l regeneration system to exam ine its
expression pattern. As shown in F ig. 5, a 900 bp
fragment corresponding toHoMADS 2 mRNA was
first ly detectab le in cu ltured exp lants at day one.
Then, theHoMADS 2 mRNA was accumulated in
flora l prmi ord ia and the regenerated flowers w ith
tepa ls at h igh leve l hormones suggesting it is re-
Fig. 4　Express ion ofHoMADS 2 in planta
HoMADS 2 mRNA was present in all floral organs in both
young f lowers and mature flow ers. No s ignal was detec ted in
vegeta tive organs inc lud ing roots, scale leaves and leaves.
HoMADS 2 mRNA accum ulation was analyzed us ing theHo-
MADS 2 3′end as the tem plate. Each lane w as loaded w ith
20μg to tal RNA. R:Roo ts;SL:Sca le leaves;L: leaves;
T1:Tepals of young f low ers;S1:Stamens of young flow-
ers;C1:Carpe ls of young f lowers;T2:Tepa ls of flowers at
anthes is;S2:S tam ens of flow ers at anthes is;C2: Carpels
of flow ers at anthes is.
quired for tepa l d ifferent ia tion. In the d ifferentia ted
stamens induced by low leve l hormones, theHoM-
ADS 2 mRNA was also detected. Th is resu lts sug-
gested that p lant hormones m ight have no effect
on the transcrip t ion ofHoMADS 2 during in vitro
flower deve lopmen.t
Fig. 5　Express ion o fHoMADS 2
in regenera ted f lowers
HoMADS 2 mRNA was detec tab le in the cu ltured explants,
the regenera ted f lowers at high leve l hormones and the d if-
ferentiated s tamens at low level hormones. RNA gel blots
(20μg) were probed w ithHoMADS 2 3′end fragm ent as the
temp late. E1:Exp lants;E2:Explants at day 5 after cultured
in the med ia w ith h igh levels of hormones;FP:F lora l prmi or-
d ia;TD1:Regenera ted flow ersw ith the firstw horl of tepa ls;
TD2:F lowers w ith a few w horls of tepa ls;SD1:S tamen pri-
mordia;SD2:Young stamens;SD3:Mature s tam ens.
3　D iscussion
In th is study, a MADS box gene,HoMADS 2,
was iso la ted from the Hyacin thus flower. Se-
1196 遗传学报　Acta Gene tica S in ica　Vo.l 32　No. 11　2005
quence comparison and phy logenetic ana lysis re-
vea led that HoMADS 2 is close ly re la ted to B
group MADS box genes and is like ly a member of
theP I fam ily instead ofAP 3 fam ily. The assump-
tion was also supported by the presence of spec if-
ic d iagnost ic sites of P I homologs in K box such
as the sequence KHENL, G lu-97 and G ly-131, at
which AP3 and PI homologs can be distingu ished
from another one (F ig. 2). In add ition, a consen-
sus sequence (MALRVQPVQPNLQE) in the
C-term ina l reg ion ofHoMADS 2 wasmore like the
P Imot if(conserved in P I fam ily genes) than the
P I-der ived motif ( conserved in AP3 fam ily
genes)[ 8, 11, 29] . Th is suggested thatHoMADS 2 is
like ly aP I homolog in Hyacin thus.
Unlike the report that the transcrip t ion ofHo-
MADS 2 fam ily genes of d icots are lmi ited in the
second and th ird whorl w ith the flower deve lop-
ment, HoMADS 2 mRNA persists in a ll flo ra l or-
gans unt il the completion of flower deve lop-
ment
[ 4, 10 , 30]
. One possib le exp lanation is that the
d ifference of express ion patterns betweenHoM-
ADS 2 and otherP I homologs in d icotsmay reflect
the d iversity o f functions inP I gene fam ily in d iffe r-
ent p lant spec ies dur ing evo lu tion. A lternative ly,
HoMADS 2 act iv ity is poss ib ly post-transcr ip iona l
regu la ted, just as L ily MADS 1 (LMADS 1), a B
group gene. LMADS 1 transcrip ts were a lso detec-
ted in a ll four f lo ra l organs and more strong ly in
la te-than in early-develop ing carpe ls. However,
LMADS1 prote inswere detected only in the peta l
and stamen, suggesting that it may funct ion v ia
post-transc rip tiona l regu la tion in peta l and stamen
development
[ 31]
.
To investigate the ro le ofHoMADS 2, we tried
to in troduce the sense HoMADS 2 driven by
CaMV 35S promoter in toA rab idopsis ecotype Co-
lumbia. However, no morpholog ica l a ltera tions
were observed in a ll transgen ic p lants (data not
shown). One explanation for these normal trans-
genic p lantsmay be that HoMADS2, a P I homolog
from hetero logous p lants, is not sufficient to form a
heterodmi er w ith A rab idopsis AP3. A lternative ly,
HoMADS2-AP3 heterodmi ers may not have the
fu ll funct ion of AP3-P I heterodmi er, which was
shown to be required for the peta l and stamen de-
ve lopment
[ 3, 4, 7]
. The smi ilar resu ltswere observed
in transgenic tobacco w ith ectop ica lly expressed
OsMADS 2, a P I homolog in rice[ 26 ] .
Prev iously, we exam ined hormone-regu la ted
in v itro flower deve lopment in hyac inthus. Further-
more, our stud ies showed that the express ions of
bothHAG 1 andHoMADS 1 were regu lated by cy-
tokin in and aux in
[ 16, 19 -21, 24]
. D iffe rent from those
found inHAG 1 andHoMADS 1, there is no obvi-
ous changes at the transcrip tion leve l o fHoMADS
2 among the exp lants, the regenerated flowers at
h igh hormone leve ls and that a t low hormone lev-
els. The data suggested that theHoMADS 2 gene
is not respons ive to cytok in in and aux in during the
regenerated flower deve lopment, while itm ight be
requ ired for tepa l and stamen differentia tion in
v itro , just as in p lanta.
In conc lus ion, we presented the character iza-
t ion of a MADS box gene, HoMADS 2, from hya-
c inthus. HoMADS 2 is expressed in a ll f lo ra l or-
gans but absent in vegeta tive organs, which was
d ifferent from that o fP I homologs in d icots. The
express ion pattern ofHoMADS 2 in the regenera-
ted flowers ind icated that its express ion is not in-
duced by the cytok in in and aux in in regenerated
flowers.
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1198 遗传学报　Acta Gene tica S in ica　Vo.l 32　No. 11　2005