全 文 ：Morphological and Cytogenetic Analysis on the
Advanced Generations of Generic Hybrids
between Brassica napus and Orychophragmus
violaceu
Zhigang ZHAO1, 2, Dezhi DU1, Zaiyun LI2*
1. National Key Laboratory Breeding Base for Innovation and Utilization of Plateau Crop Germplasm of Qinghai Province, Acade-
my of Agriculture and Forestry, Qinghai University，Xining 8100161, China;
2. National Key Lab of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China
Supported by the Special Fund for Preliminary Study of the National Basic Research
Program of China (973 Program， 2011CB111508).
*Corresponding author. E-mail: 13897474887@126.com
Received: March 10, 2012 Accepted: June 26, 2012A
Agricultural Science & Technology, 2012, 13(7): 1410-1414，1446
Copyright訫 2012, Information Institute of HAAS. All rights reserved Agricultural Biotechnology
Abstract [Objective] This study aimed to reveal the genetic changes of advanced
generation hybrids between Brassica napus and Orychophragmus violaceus. [Method]
The morphological characteristics such as the plant shape, branching sites, leaf
shape, leaf color, primary branches and secondary branches, as well as the cytolog-
ical characteristics of the advanced generation hybrids (F8-F10) between B. napus
and O. violaceus were observed. [Result] The morphology analysis revealed that the
hybrid progeny was more like B. rapa in leaf shape, leaf color, plant shape and ear-
ly flowering phenotype, whereas more like B. napus in number of secondary branch-
es, silique length and 1 000-seed weight. Analysis on the cytogenetics characteristics
showed that these advanced inbred progenies were hypoploids with less than 38
chromosomes; moreover, all the chromosomes from O. violaceus had been lost.
Chromosome pairings at meiotic diakinesis of hybrids between the advanced inbred
progenies and B. rapa revealed that chromosomes lost in hypoploids possibly be-
longed to the C genome of B. oleracea. With generations developing, chromosomes
number of plants from two populations gradually increased and developed into the
number of B. napus (2n=38). [Conclusion] This study will provide reference to reveal
the source of chromosome lost in hypoploids and the morphological change of
hybrids.
Key words Brassica napus; Orychophragmus violaceus; Sexual hybrids; Hypoploid
A llopolyploids are developedsuccessfully in nature, whilethe synthetic polyploids are
usually instable in genetic and mor-
phological aspects [1]. Such instability
refers to the nondirectional change of
the phenotype and genomic structure,
which may be resulted from the
changes in chromosome number and
structure, such as chromosome elimi-
nation, doubling, rearrangement, and
epigenetic modification of DNA, which
may be caused by the distant parental
genetic relationship. It is speculated
that during the polyploid formation in
nature, such instability is eliminated by
the adaptability in evolution to produce
the stable species. In the synthetic
distant hybrids, several cytological
abnormalities were often found, for ex-
ample, the chromosome elimination
and mitosis are not synchronized [2-4].
These cytological abnormalities can be
tentatively explained by the instability
of the distant hybrids, but its molecular
mechanism is still unclear.
Orychophragmus violaceus (OE
Schulz, 2n=24), also known as the Er-
yuelan, belongs to the Orychophrog-
mus in Brassica, Cruciferae. In this
study, the morphological and cytologi-
cal characteristics of the advanced
generation hypoploid hybrids between
B. napus and O. violaceus were ob-
served to reveal the reasons causing
the loss of chromosome in hybrid hy-
poploid and morphological changes.
Materials and Methods
Materials
Li et al. obtained the intergeneric
mixoploid hybrid of Brassica napus
(2n=38) cultivar Oro and Orychophra-
gmus violaceus by sexual hybridiza-
tion[5]. The hybrid has certain fertility as
its parental chromosomes are com-
pletely or partly separated in mitosis
and meiosis. After successive selfing,
one plant with yellow flowers was
found in the 5th generation (F5), whose
chromosome behavior was different
from the performance of earlier gener-
ations[6-7]. This plant with yellow flower
in F5 was selfed to produce 18 different
offspring plants (F6). Among the 18
plants, three were more like B. rapa, in
which, two plants were completely
sterile, another plant was selfed to pro-
duce the F7. From the F7 plants, two
plants which were obviously like B. ra-
pa were selected and selfed to pro-
duce the F8, F9 and F10, which were di-
vided into linesⅠand II.
Methods
Morphological observation First of
all, 15 -30 mature plants were ran-
domly selected from the linesⅠandⅡ,
the three control varieties (two B. rapa
cultivars Hongqiaoaiqingcai and Qing-
you241, B. napus Oro) and then dried.
Their agronomic traits including the
plant height, branching sites, number
of primary branches, number of sec-
ondary branches, length of the main
inflorescence, silique number of the
main inflorescence, sillique number
DOI:10.16175/j.cnki.1009-4229.2012.07.025
Agricultural Science & Technology
Vol.13, No.7, 2012 Agricultural Science & Technology
2012
Plant
length
cm
Branch
position
cm
No. of
primary
branches
No. of
second
branches
Length of
main
inflorescen-
ce∥cm
No. of
siliques on
main
inflorescence
Silique
length
cm
No. of
seeds in
silique
No. of silique 1 000 seedsweight∥g
125.4±13.1 25.0±9.9 5.2±2.3 4.6±2.6 71.9±13.9 57.4±12.8 6.4±0.4 22.3±3.5 215.1±71.7 3.8±0.5
(108.1-149.7) (9.1-37.2) (2-9) (0-9) (48.2-90.1) (36-87) (5.6-7.1) (16.1-30.3) (105-345) (3.1-4.8)
123.6±12.1 19.8±11.3 5.1±1.2 5.2±2.2 69.0±12.1 51.6±12.3 6.4±0.7 23.4±4.8 240.9±80.4 3.6±0.6
(103.9-150.2) (5.1-54.1) (3-7) (1-9) (47.0-90.2) (31-78) (5-7.8) (16.2-30.6) (113-398) (2.3-4.6)
136.5±11.1 16.2±10.4 5.6±2.2 7.4±3.4 76.5±9.1 55.3±9.3 4.9±0.4 23.38±3.1 281.3±93.2 2.5±0.3
(118.1-158.2) (0-33.2) (2-10) (2-14) (56.0-90.1) (42-74) (4.0-5.8) (16.6-29.0) (140-449) (2.0-3.1)
140.1±11.2 14.3±10.4 5.8±1.1 8.1±3.3 67.8±13.9 56.4±10.9 5.4±0.8 19.8±3.2 376.0±99.3 2.9±0.5
(127.1-160.0) (0-31.7) (4-8) (3-13) (48.2-70.8) (43-71) (4.8-7.4) (14.0-23.8) (236-497) (2.1-3.7)
173.1±13.1 51.85±19.9 6.4±1.8 5.4±4.1 79.6±14.4 65.2±16.6 6.4±0.7 22.9±2.9 357.9±132.1 3.7±0.5
(144.1-194.2) (21.0-80.9) (4-10) (2-14) (50.1-99.7) (34-95) (5.0-7.4) (19.8-28.6) (141-581) (3.1-5.1)
Table 1 Botanical characters of two F10 lines of B. rapa-like progenies
Group
Ⅰ
Ⅱ
Hongqiao
aiqingcai
Qingy-
ou241
Oro
of the whole plant, length of the
silique, grain number in a silique and
1 000-seed weight were measured.
Cytological observation Treat-
ments on ovary and anther, as well as
the cytological observation were per-
formed according to the method by Li
et al. [5] In order to clear the chromo-
somes lost in the hypoploids were
from genome A or C, the hypoploids of
F9 and F10 which were more like B. ra-
pa were crossed with B. rapa plants to
produce the hybrids, of which, the
chromosome pairing in meiotic diaki-
nesis were observed.
Genomic in situ hybridization The
genomic in situ hybridization was con-
ducted according to the method by
Zhong et al. [8 -9], with some modifica-
tions. In this study, the O. violaceus
genome was labeled as a probe by
nick translation, and the B. napus Oro
genome was the blocking DNA.
Results and Analysis
Morphological characteristics of
the advanced generation hybrids
Two plants of F7 (1-8 Yi and 2-5
Han) were more like B. rapa in leaf
type, leaf color, plant type, earliness,
etc. The segregation of character
within or between linesⅠ and Ⅱ was
observed in the plants of F8-F10. The
traits segregation within lines was
mainly reflected in the growth potential
of the plant (such as plant height and
branch number), as well as in leaf type
and leaf color. For example, in the F9
line II, some plants had bright green
leaves with little or no wax on surface,
more similar to B. rapa, while other
plants had fat leaves covered with
thick layer of wax, more similar to
B. napus.
In the hybrid offsprings, the three
traits including early flowering, small
plants and similarity to B. rapa were
correlated, namely, the B. rapa-like
hybrids were generally small and flow-
ered earlier. The investigation on the
two lines of F9 in 2005 in Qinghai re-
vealed that most plants in the lines I
and II flowered earlier. The B. rapa-like
hybrids and the two B. rapa cultivars
(Hongqiaoaiqingcai and Qingyou241)
flowered synchronously, 10-15 d earli-
er than B. napus. The hybrids matured
3-5 d later than B. rapa cultivars but
9 -15 d earlier than B. napus . The
B. rapa-like hybrids sowed in Wuhan
flowered especially earlier. The hybrid
planted in October, 2005 flowered in
the late September, and the parent
plants OrO did not flower till the next
late February or early March.
The hybrid progeny was more like
B. rapa in leaf shape, leaf color and
growth period (Fig.2). In addition, the
hybrid plant was shorter, like B. rapa,
but significantly lower than Oro. The
branching site of the hybrid was more
like B. rapa plants. The plants of lines
Ⅰ and II showed no significant differ-
ences with B. rapa and B. napus in
number of primary branches, main in-
florescence length and the silique
number of the main inflorescence
(Table 1).
The hybrid progeny was more like
B. napus Oro in number of secondary
branches, silique length and 1 000 -
seed weight, significantly different from
B. rapa. The average grain number of
a silique was similar in the lines I and
II, B. rapa and Oro; the silique number
of whole plant of lines I and II was sig-
nificantly less than that of other culti-
vars (Table 1).
Cytological characteristics
Genomic in situ hybridization (GISH)
analysis Five plants were selected
from each of the lines I and II in F9, and
the chromosome of pollen mother cells
(PMCs) in meiotic diakinesis and
anaphase I was investigated using
GISH technique. No signal from the
complete chromosome or chromoso-
mal fragment of O. violaceus was de-
tected in all the tested PMCs. The red
probe signal was concentrated in the
ends and the centromeric region ofFig.1 The sources and pedigree of the experimental materials
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Agricultural Science & Technology Vol.13, No.7, 2012
2012
(A) young plants from lineⅠof B. rapa-like F9 progenies; (B) young plants from line Ⅱof
B. rapa-like F9 progenies; (C) one ovary cell (2n = 34) at diakinesis (17 Ⅱ +1 Ⅰ ); (D) a
PMC at diakinesis (17Ⅱ +1Ⅰ), the arrowhead shows a univalent, (bar: 10 μm) ; (E) mei-
otic diakinesis (10Ⅱ +7Ⅰ ) of a hybrid (2n = 27) between a plant from F8 lineⅡand B. ra-
pa, the arrowheads show univalents (bar: 10 μm); (F) emerged image of a diakinesis PMC
of a plant from F9 lineⅠ, red signals from labeled O. violaceus probe were mainly located
on terminal or centromeric region of some chromosomes (bar: 10 μm).
Fig.2 Morphology and cytology of B. rapa-like progenies of sexual hybrids between B. na-
pus and O. violaceus
some chromosomes (Fig.2 -F), sug-
gesting that the O. violaceus chromo-
somes had been lost in these plants.
Somatic chromosomenumber Am-
ong the tested nine plants of line Ⅰ in
F8, four were non-mixoploid (all somat-
ic cells have the same number of
chromosomes), containing 34 chro-
mosomes; and the other five were
mixoploid (the chromosome number is
not exactly the same in all the somatic
cells), among them there were two
plants with the highest frequency of 34
chromosomes, (Fig.2-C), one with the
highest frequency of 33 chromo-
somes, one with the highest frequency
of 36 chromosomes and one with
highest frequency of 37 chromo-
somes. Among the tested 21 plants of
line Ⅰ in F9, six were non-mixoploid
(three had 33 chromosomes, two had
36 chromosomes and one had 37
chromosomes); the other 15 were
mixoploid, among them, there were
three plants with the highest frequency
of 34 chromosomes, one plant with the
highest frequency of 35 chromo-
somes, three plants with the highest
frequency of 36 chromosomes, two
plants with the highest frequency of 37
chromosomes and one plant with the
highest frequency of 38 chromo-
somes. All the three tested plants of
line II in F8 were mixoploid, including
one plant had the highest frequency of
34 chromosomes and two plants had
the highest frequency of 36 chromo-
somes. Among the tested 13 plants of
line Ⅱ in F9, five were non-mixoploid
(two had 34 chromosomes, two had 38
chromosomes and one had 36 chro-
mosomes). Among the tested 27
plants of line Ⅱ in F10, five were non-
mixoploid (one had 36 chromosomes,
two had 37 chromosomes and one
had 38 chromosomes); among the 22
mixoploids, two plant had the highest
frequency of 34 chromosomes, seven
had the highest frequency of 35 chro-
mosomes, eight had the highest fre-
quency of 36 chromosomes, two had
the highest frequency of 37 chromo-
somes and three had the highest fre-
quency of 38 chromosomes.
Table 2 shows that among all the
detected cells of lineⅠ in F8, the cells
with chromosomes of 2n = 34 were the
most, accounting for 36.6% , followed
by cells with chromosomes of 2n = 36,
accounting for 23.9% , the cells with
chromosomes of 2n = 38, accounting
for 1.4 % only. In F9 generation, the
cells with chromosomes of 2n = 36
were the most frequent, accounting for
36.7% , and the cells with chromo-
somes of 2n = 34 dropped to 14.4% ;
the cells with chromosomes of 2n = 38
increased from 1.4% in F8 to 9.3% .
Among the cells of lineⅡ in F8, F9 and
F10, the cells with chromosomes of 2n=
36 were the most frequent, accounting
for 36.8% , 31.7% and 30.5% in the
three generations respectively. The
cells with the chromosomes of 2n> 36
were increased from F8 to F10, which is
similar in the linesⅠandⅡ (Table 2).
Meiosis The linesⅠ and II in differ-
ent generations mainly produced chro-
mosome in the forms of 17 II, 18 II, 19
II, 16 II +2Ⅰ or 3Ⅰ, 17 II +1Ⅰ or 2Ⅰ,
18 II +1Ⅰ , etc. in meiosis diakinesis
(Fig. 2D). The pollen mother cells con-
taining 16 bivalents were found only in
one plant of lineⅠin F9.
In all the detected plants, the nu-
clei in meiosis anaphase I contained
15, 16, 17, 18, 19 or 20 chromosomes
(Table 3). The nucleus containing 18
chromosomes was the most frequent
in F9 line I, accounting for 37.5%, fol-
lowed by the nuclei containing 17
chromosomes, accounting for 25%; in
14.3 % of the PMCs at anaphase I,
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2012
Table 2 Somatic chromosome numbers of two lines of B. rapa-like progenies
Lines Generation No.ofplants
Number and percentage of somatic cells with chromosome number∥% Total number
of cells≤30 31 32 33 34 35 36 37 38
Ⅰ F8 9 2(2.8) 1(1.4) 4(5.6) 4(5.6) 26(36.6) 10(14.1) 17(23.9) 6(8.5) 1(1.4) 71
F9 21 1(0.5) 3(1.5) 6(3) 3(1.5) 31(14.4) 14(6.5) 79(36.7) 58(27) 20(9.3) 215
Ⅱ F8 3 2(10.5) 3(15.8) 4(21.1) 7(36.8) 3(15.8) 19
F9 13 1(1.7) 5(8.3) 3(5) 12(20) 3(5) 19(31.7) 3(5) 14(23.3) 60
F10 27 5(2.2) 4(1.7) 7(3.0) 23(9.9) 51(21.9) 71(30.5) 39(16.7) 33(14.2) 233
Data out and in the brackets are number and percentage of cells, respectively.
Table 3 Chromosome segregations in AⅠPMCs of two lines of B. rapa-like progenies
Lines Gener-ations
Chromosome numbers in each polar group∥% PMCs with
laggards∥%15 16 17 18 19 20
Ⅰ F9 12.5 25.0 37.5 16.7 8.3 14.3
Ⅱ F9 1.8 7.3 40.0 16.4 30.9 3.6 17.8
F10 2.6 18.4 31.6 36.8 10.5 40.7
lagging chromosomes were observed.
In line Ⅱ of F9, the nuclei with 17
chromosomes were the most frequent
at anaphase I, accounting for 40% ,
followed by the nuclei with 19 chromo-
somes, accounting for 30.9%; the nu-
clei with 18 chromosomes were the
third most frequent, accounting for
16.4%. In the lineⅡ, lagging chromo-
somes were observed in 17.8% of the
PMCs at anaphase I. In F10, nuclei at
anaphase I containing 19 chromo-
somes were the most frequent, ac-
counting for 36.8% , and the nuclei
containing 18 chromosomes were in-
creased to 31.6%, and the nuclei con-
taining 17 chromosomes accounted
for only 18.4%. Lagging chromosomes
were observed in 40.7% of the PMCs
of F10 at anaphase I.
Meiosis of hybrids between the ad-
vanced inbred progenies and B. ra-
pa By counting the chromosomes in
somatic cells, we selected an F1 plant
that had 27 chromosomes and an F1
plant that had 28 chromosomes to ob-
serve their chromosome pairing during
meiotic phase. We found that most of
the pollen mother cells (PMCs) at di-
akinesis contained chromosomes in
form of 10 II +7 I, and less PMCs had
the chromosomes in form of 11 II +6
Ⅰin the F1 plant of 2n = 27 (Fig. 2E).
Most of the PMCs with 2n = 28 at di-
akinesis contained chromosomes in
form of 10 II +8Ⅰ.
Discussions
Only one plant with the pale yel-
low flowers were found in the F5 gen-
eration of the distant hybrids between
B. napus and O. violaceus, while all
the plants in the early generations had
yellow flowers, which were same to
the female parent B. napus. The pollen
mother cells of this plant had 31 chro-
mosomes, which were separated as
15:16 or 12:19 in meiotic anaphase I.
The separation of 15:16 may be de-
rived from the separation of A and C
genomes in hybrid cells, which was
partly proved in phenotype by the oc-
currence of B. rapa-like progeny. B.
napus (AACC) was developed from
hybrid between B. rapa (AA) and B. ol-
eracea (CC) through chromosome
doubling [10]. As the characteristics of
original B. rapa were unclear, the sep-
aration of A and C genomes may dif-
ferentiate progeny genetically close to
the original B. rapa parent. The initial
purpose of this study was to screen
the plants most similar to B. rapa par-
ent in phenotype from the B. rapa-like
progeny and to further determine their
genome compositions, so that it can
provide foundation for research on
evolution of B. napus. However, we
found that even the plants most like
B. rapa in phenotype, they still had
several indices (number of secondary
branches, silique length, 1 000-seed
weight etc.) closer to the those of B.
napus. Because the plants were se-
lected according to their leaf color, leaf
type, plant type, flowering time and
other intuitive morphological indices,
the quantitative traits such as 1 000-
seed weight were not considered, the
B. rapa- like plants were similar to
B . rapa just in phenotype to some
extent.
We initially speculated that the
reason for the differentiation of the
B. napus-like progeny may be that the
C chromosomes from female parent
were partly or completely eliminated,
which gave more space to express the
A genome. By counting chromo-
somes, we found that in all the cells
observed in the F8 and F9 generations
of lines I and Ⅱ , the cells containing
chromosomes of 2n ≤ 30 shared a
very small proportion, no more than
2.8%. Most aneuploid cells contained
34-37 chromosomes, suggesting that
1-4 parental chromosomes were lost
frequently. We observed the chromo-
some pairing in PMCs at meiotic
phase to reveal the source of the lost
chromosome. And we found that most
of the pollen mother cells (PMCs) at
diakinesis contained chromosomes in
form of 10 II +7 I, and less PMCs had
the chromosomes in form of 11 II +6
Ⅰ in the F1 plant of 2n = 27 (Fig.2E).
Most of the PMCs with 2n = 28 at di-
akinesis contained chromosomes in
form of 10 II +8 Ⅰ . Leflon et al. found
that at the meiotic diakinesis of AAC
triploid, only 0.2 C genome homolo-
gous bivalent could be detected in a
PMC on avarage, while the A genome
could form 10 bivalents[11]. In the F1 hy-
brids between the advanced inbred
progenies and B. rapa, most PMCs
had 10 bivalents at the meiotic diakine-
sis, it was probably because that the
10 A chromosomes from B. rapa
matched preferentially with 10 A chro-
mosomes from the hybrids, while the
C chromosomes formed univalents.
We speculated the C chromosomes
rather than A chromosomes were lost
in the B. rapa-like progeny.
With generations developing,
chromosomes number of plants from
two populations gradually increased
and developed into the number of B.
napus (2n=38). In the lineⅠ, the cells
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Agricultural Science & Technology Vol.13, No.7, 2012
2012
(Continued on page 1446)
Responsible editor: Qingqing YIN Responsible proofreader: Xiaoyan WU
with chromosomes of 2n = 38 from F8
to F9 were increased from 1.4% in F8 to
9.3% in F9, while the cells with chro-
mosomes of 2n > 34 were increased
from 48.0% to 80.0% . In line Ⅱ , the
cells with chromosomes of 2n = 38
were gradually increased from 15.8%
in F8 to 30.9% in F10. All the detected
cells with chromosomes of 2n < 38
were hypoploids. Tokumasu obtained
a hypoploid of 2n =32 through parth-
enogenesis from a polyploidy line of
B. napus. This hypoploid produced
gametes which showed different vital-
ities. Among them the gamete with
nearly 19 chromosomes exhibited high
vitality, and the gamete with 19 chro-
mosomes had the highest vitality; in
addition, the number of chromosomes
of its selfed progeny ranged from 2n =
33 to 2n = 38, and the progeny with
2n = 38 was the most. He also ob-
served that the fertility of plants in-
creased with the increase of chromo-
some number[12]. Similar situation also
occurred in hyperploid, that the chro-
mosome number is prone to the num-
ber in original parent. Tokumasu found
that chromosome number of the selfed
progeny was lower than their parental
hyperploid containing 76, 67, 58, 52
and 46 chromosomes, reduced in a
trend down to 2n = 38[12]. Among the
progeny of the pentaploid hybrid be-
tween B. napus and O. violaceus pen-
taploid, the aneuploid cells or pollen
mother cells with 47, 44 and 41 chro-
mosomes were more frequent than
other cells. The chromosome reduc-
tion were observed in the progeny of
cells with 2n=44 and 2n=4, and chro-
mosomes were reduced faster in male
gametes, while the more gametes with
less chromosomes (n=19-22) passed
to the future generations[13]. Similar ph-
enomenon was observed in the diploid
offspring between B. japonica and
Raphanus sativus[14].
The allopolyploid flowering plants
probably developed from diploid
species through sudden and unex-
pected polyploidization and then grad-
ual diploidization[15-16]. B. napus (AACC,
2n =38) after long-term evolution has
formed stable diploidized allote-
traploid, while other allopolyploids and
hyperploids may be eliminated by na-
ture or human because of their infertil-
ity and poor vitality. In this study, the
advanced generation hybrids between
B. napus and O. violaceus were ana-
lyzed. Cytological analysis indicated
that these hybrid progenies were hy-
poploids and most C chromosome
were lost, that is the main reason why
the hybrids was more like B. rape in
phenotype. Based on these results, we
could predict that the plant morphology
may change reversibly from B. rapa-
like to B. napus-like as the generation
advances and the number of chromo-
somes gradually returns to 2n=38.
References
[1] COMAI L. Genetic and epigenetic inter-
actions in allopolyploid plants[J]. Plant
Mol Biol, 2000, 43: 387-399.
[2] RIERA-LIZARAZU O, RINES HW, PH-
ILLIPS RL. Cytological and molecular
characterization of oat × maize partial
hybrids[J]. Theor Appl Genet, 1996, 93:
123-135.
[3] IWAMATSU T, KOBAYASHI H, YA-
MASHITA M, et al. Experimental hy-
bridization among Oryzias species. II.
Karyogamy and abnormality of chromo-
some separation in the cleavage of in-
terspecific hybrid between Oryzias
latipes and O. javanicus [J]. Zool Sci,
2003, 20: 1381-1387.
[4] BARBOSA LV, VIEIRA MLC. Meiotic
behavior of passion fruit somatic hy-
brids, Passiflora edulis f. flavicarpa
Degener + P. amethystina Mikan [J].
Euphytica, 1997, 98: 121-127.
[5] LI Z, LIU HL, LUO P. Production and
cytogenetics of intergeneric hybrids be-
tween Brassica napus and Orychophra-
gmus violaceus [J]. Theor Appl Genet,
1995, 91: 131-136.
[6] LI ZY (李再云 ). The chromosome be-
havior and evolution significance of
Brassica napus L. and Orychophrag-
mus violaceus new hybrid material(甘蓝
型油菜与诸葛菜属间杂交新材料的染色
体行为及其进化意义 ) [J]. Progress in
Natural Science (自然科学进展), 2003,
13: 807-813.
[7] ZHAO ZG, MA N, LI ZY. Alteration of
chromosome behavior and synchro-
nization of parental chromosomes after
successive generations in Brassica na-
pus × Orychophragmus violaceus hy-
brids[J]. Genome, 2007, 50: 226-233.
[8] ZHONG XB, HANS DE JONG J, ZABEL
P. Preparation of tomato meiotic
pachytene and mitotic metaphase chro-
mosomes suitable for fluorescence in
situ hybridization (FISH) [J]. Chromo-
some Res, 1996, 4: 24-28.
[9] LEITCH AR, SCHWARZACHER T,
JACKSON D, et al. Microscopy Hand-
book No.27. In situ hybridization: a
practical guide[M]. Oxford: Bios Scien-
tific, 1994.
[10] UN. Genomic analysis in Brassica with
special reference to the experimental
formation of B. napus and peculiar
mode of fertility [J]. Japan J Bot, 1935,
7: 389-450.
[11] LEFLON M, EBER F, LETANNEUR
JC. Pairing and recombination at
meiosis of Brassica rapa (AA) × Bras-
sica napus (AACC) hybrids [J]. Theor
Appl Genet, 2006, 113: 1467-1480.
[12] TOKUMASU S. A hypodiploid of rape
(Brassica napus L.) and chromosome
variation in its progeny[J]. Jpn J Genet,
1984, 59: 527-535.
[13] WU JG, SHI CH, LI ZY, et al. Analysis
of cytogenetics for the aneuploids in
pentaploid progenies from intergeneric
cross between Brassica napus and
Orychophragmus violaceus[J]. J Zhe-
jiang Univ, 2002, 28: 601-608.
[14] KATO M, TOKUMASU S. The stabi-
lization of chromosome numbers and
the maintenance of euploidy in Brassi-
coraphanus [J]. Euphytica, 1983, 32:
415-423.
[15] WOLFE KH, SHIELDS DC. Molecular
evidence for an ancient duplication of
the entire yeast genome [J]. Nature,
1997, 387: 708-713.
[16] SOLTIS D, SOLTIS P, TATE J. Ad-
vances in the study of polyploidy since
plant speciation[J]. New Phytol, 2003,
161: 173-191.
[17] ZHAO FY (赵福永), NU JX (鲁军雄),
HUANG J (黄杰) .Genetic analysis on
interspecific hybrids of brassica napus
and wild brassica juncea(甘蓝型油菜与
野生芥菜型油菜杂交子代的遗传分析)
[J]. Journal of Anhui Agricultural Sci-
ences (安徽农业科学 ), 2011,39 (2):
718-720, 794.
[18] SHI JF, YIN CY, RONG L, et al. Het-
erosis analysis of nitrogen use efficien-
cy for grain production of Brassica na-
pus L.[J]. Agricultural Science & Tech-
nology, 2010, 11(1):45-47.
[19] HE JP(何佳平), ZHANG CH(张从合 ),
JIANG L (江力 ), et al. Study on the
quality characteristics of hybrid rape
cultivars and their parents(杂交油菜品
种及其亲本品质特性研究 ) [J]. Jour-
nal of Anhui Agricultural Sciences (安
徽农业科学), 2010,38(2):657-658.
[20] HUANG ZS, PAISAN L, THITIPORN
M, et al. Correlation analysis on male
sterile lines characters in rapeseed
(Brassica napus L.)[J]. Agricultural Sci-
ence & Technology, 2010, 11 (9-10):
183-187.
1414
Agricultural Science & Technology
Agricultural Science & Technology Vol.13, No.7, 2012
2012
(Continued from page 1414)
甘蓝型油菜和诸葛菜属间杂种高世代材料形态学和细胞学分析
赵志刚 1，2，杜德志 1，李再云 2* （1.青海大学农林科学院青海省高原作物种质资源创新与利用国家重点实验室培育基地，青海西宁 810016；2.华
中农业大学作物遗传改良国家重点实验室，湖北武汉 430070）
摘 要 [目的]进一步揭示甘蓝型油菜和诸葛菜属间远缘杂种高世代（F8~F10）材料的遗传变异规律。[方法]以甘蓝型油菜和诸葛菜属间杂种高世
代群体为材料，对甘蓝型油菜和诸葛菜远缘杂交后代表型偏白菜型油菜变异群体进行形态、细胞学方面的研究。[结果]形态学分析表明，杂种后
代叶型、叶色、株型、早花性状偏向白菜型油菜，而二级分枝数、角果长度和千粒重与甘蓝型油菜 Oro更为接近；细胞学研究表明，这些杂种高世代
材料细胞中已不含诸葛菜染色体，多数植株都为少于 38条染色体的亚倍体；与白菜型油菜杂种的减数分裂配对结果显示，这些亚倍体后代细胞
中丢失的可能是来自 C基因组的染色体，随着世代的增加，体细胞染色体数目都有向甘蓝型油菜 2n=38升高和回归的趋势。[结论]该研究为揭示
杂种后代亚倍体中丢失染色体的来源及形态学改变的原因奠定基础。
关键词 甘蓝型油菜；诸葛菜；有性杂种；亚倍体
基金项目 “973”前期研究（2011CB111508）。
作者简介 赵志刚（1978-），男，内蒙古多伦县人，副研究员，博士，硕士生导师，从事油菜细胞遗传学研究，E-mail: 13897474887@126.com。*通讯作者。
收稿日期 2012-03-10 修回日期 2012-06-26
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
能源植物潜力种——甘肃大戟（Euphorbia kansuensis Prokh.）的形态和分布研究
顾子霞，郭建林，周义锋，杭悦宇 * （中国科学院植物研究所，江苏省植物迁地保护重点实验室，江苏南京 210014）
摘 要 [目的]研究甘肃大戟的分类学地位。[方法]根据野外调查、采集，结合标本观察，对江苏分布的乳浆大戟亚属的甘肃大戟（Euphorbia
kansuensis Prokh.）和月腺大戟（E. ebraceolata Hayata）的形态特征和分布区等方面进行了研究。[结果]《江苏植物志》记载的月腺大戟和《中国植物
志》的甘肃大戟在形态描述的被毛和根形态这 2个重要特征上没有明显区别，且相互有交叉，故《中国植物志》认为月腺大戟是甘肃大戟的误用名
是合理的，但是其形态描述应修订为：全株有毛或无毛；根为纺锤形至圆柱状；腺体为肾状长圆形。[结论]该研究对于理清甘肃大戟的分类学地位
具有重要意义。
关键词 能源植物；月腺大戟；甘肃大戟；形态；分布
基金项目 江苏省农业科技攻关重点项目(BE2008328)。。
作者简介 顾子霞（1985-），女，江苏常熟人，硕士，从事能源植物研究，E-mail:zixia_16@163.com。*通讯作者，研究员，博士，博士生导师，从事植
物多样性与系统演化研究，E-mail:hangyueyu@yahoo.com.cn。
收稿日期 2012-03-08 修回日期 2012-06-28

Responsible editor: Yongbo DUAN Responsible proofreader: Xiaoyan WU
and concentrated in Gansu, Qinghai
and Sichuan, with a diminishing ten-
dency from West to East according to
Flora of China and its English version.
Taking the sites of collection of speci-
mens and the modified classification of
E. ebraceolata Hayata into considera-
tion, Anhui and Tibet should be added
to the distribution areas of E. kan-
suensis Prokh.
References
[1] China Plant Flora Compiling Committee,
Chinese Academy of Science(中国科学
院中国植物志编辑委员会). China plant
flora Vol. 44, Section 3(中国植物志第四
十四卷 , 第三分册)[M]. Beijing: Science
Press(北京: 科学出版社), 1997: 89-90.
[2] WU ZY, RAVEN PH, HONG DY, et al.
Flora of China (Vol.11)[M]. Beijing: Sci-
ence Press, and St. Louis: Missouri
Botanical Garden Press, 2008: 305.
[3] FU LG (傅立国). Higher plants in China
Vol. 8(中国高等植物第八卷)[M]. Qing-
dao: Qingdao Press(青岛: 青岛出版社),
2001: 191.
[4] MA JS (马金双), WU ZY (吴征镒). New
material of Euphorbia in China (国产大
戟属新资料)[J]. Acta Botanica Yunnani-
ca(云南植物研究), 1992(4): 362-272.
[5] China Oil Plant Compiling Committee(中
国油脂植物编写委员会). China oil plant
(中国油脂植物 ) [M]. Beijing: Science
Press(北京: 科学出版社), 1987.
[6] CHENG BJ(程柏君), QIAN XS(钱学射),
GU GP(顾龚平), et al. The comprehen-
sive utilization of Euphorbia lathyris and
its cultivation(能源作物续随子的综合利
用和栽培 ) [J]. Chinese Wild Plant Re-
sources (中国野生植物资源), 2007, 26
(4): 19-22.
[7] GU ZX (顾子霞), WU BC (吴宝成), WU
LY (吴林园), et al. Analysis of total lipid
contents and fatty acids composition of
three species of Euphorbia in Jiangsu
Province(江苏 3 种大戟属野生植物总脂
含量及脂肪酸组分分析)[J]. Chemistry &
Industry of Forest Products(林产化学与
工业), 2009, 29(4): 63-66.
[8] ZHAO ZL(赵志礼), ZHAO RN(赵汝能).
Examine and correct of botanical origin
of Tibetan medicine in Sichuan (藏药川
布的原植物考订)[J]. Chinese Pharma-
ceutical Journal (中国药学杂志), 1992,
27(5): 269-270.
[9] HAYATA B. Revisio Euphorbiacearum
et Buxacearum Japonicarum [J]. Journ
Coll Sci Tokyo, 1904, 3: 71.
[10] Jiangsu Institute of Botany(江苏省植物
研究所). Jiangsu plant flora (江苏植物
志 ) [M]. Nanjing: Jiangsu Science and
Technology Publishing House(南京: 江
苏科学技术出版社), 1982: 414.
[11] Anhui Plant Flora Association Group
(《安徽植物志 》 协作组 ). Anhui plant
flora Vol.3 (安徽植物志第三卷 ) [M].
Hefei: Anhui Science and Technology
Publishing House(合肥: 安徽科学技术
出版社), 1990: 257.
[12] Zhejiang Plant Flora Compiling Com-
mittee (浙江植物志编辑委员会). Zhe-
jiang plant flora Vol.3(浙江植物志第三
卷 ) [M]. Hangzhou: Zhejiang Science
and Technology Publishing House (杭
州: 浙江科学技术出版社), 1993: 491.
[13] CHEN HB (陈汉斌), ZHENG YJ (郑亦
津), LI FZ(李法曾). Shandong plant flo-
ra(山东植物志)[M]. Qingdao: Qingdao
Press(青岛: 青岛出版社), 1997: 536.
[14] ZHOU SQ(周世权), LIU GH(刘果厚). A
new material of Euphorbia in China(中
国大戟属一新种)[J]. Acta Phytotaxo-
nomica Sinica (植物分类学报), 1989,
22(1): 77-78.
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