al12 水稻苗期低温白化突变体al12的超微结构与基因定位
遗 传 学 报 Acta Genetica Sinica , December 2006, 33 (12)：1112–1119 ISSN 0379-4172
Ultrastructure and Gene Mapping of the Albino Mutant al12 in Rice (Oryza sativa L.)
XIA Jiu-Cheng 1,2,3, WANG Yu-Ping 1,2, MA Bing-Tian 1,2, YIN Zhao-Qing 1,2, HAO Ming 1,2, KONG De-Wei 1,2 , LI Shi-Gui 1,2,
①
1. Rice Institute of Sichuan Agricultural University , Wenjiang 611130, China ;
2. Key Laboratory of Crop Genetic Resources and Improvement , Ministry of Education , Sichuan Agricultural University , Ya ’an 625014, China ;
3. Panzhihua University , Panzhihua 617000, China
Abstract: Seedling albino mutation resistant to low temperature is an adaptability of rice (Oryza sativa L.) to cold. The mutant, a conditional expression controlled by development and temperature, differs from other albino mutants. The chlorophyll content of the mutant was measured using a portable chlorophyll meter, and the ultrastructure of the chloroplast was observed using a transmission electron microscope. Chlorophyll content was 1.2 SPAD, and the chloroplast did not develop, with only small vesicle-like structures. A segregation analysis of the reciprocal crosses between the albino mutation line with the rice line 9311 demonstrated that the albino trait was controlled by a single recessive gene, which was flanked by SSR markers RM5068 and RM3702 on the short arm of chromosome 8 with a distance of 0.5-1.1 cM and 4.9 cM, respectively. This gene was mapped within a 6 cM interval region and was tentatively referred to as al12. Key words: rice; albino mutant; chloroplast; gene mapping
Received: 2005-12-22; Accepted: 2006-04-03
This work was supported by Program for New Century Excellent Talents in University (No. NCET-04-0907) and Program for In-novative Research Team in University (No.IRT0453). ① Corresponding author. E-mail: Lishigui_sc@https://www.360docs.net/doc/9112069767.html,
At present, many albino mutants have been found in rice (Oryza sativa L.). Iwata et al .[1,2] and Maekawak et al .[3] reported 11 mutants, from al1 to al11. Shu et al .[4] reported the albino mutant W 25, and YU et al .[5] reported the albino mutant alb21. How-ever, most of these are lethal mutants [1,2,5], having a low value for both theoretical and practical applica-tions. A new mutant (259) was found in our labora-tory, which behaves like an albino before the 3-leaf-stage (only with endosperm supply energy) at a low temperature (<24), but after the 3℃-leaf-stage, the mutant gradually turns green and completes the reproduction. It is a conditional expression mutant and is directly related to the adaptation of the plant and can be a unique material for researching the ad-versity adaptation of rice. By crossing and back-crossing with 9311, the separation for normal and albino plants in the progenies fits well to a ratio of 3:1, suggesting that the mutant is controlled by a re-cessive gene. The gene was tentatively named al12. In this article, the chlorophyll content was calculated using a portable chlorophyll meter and the ultrastruc-ture change of the chloroplast was observed using a transmission electron microscope.
Recently, with the rapid development of the SSR molecular marker technique, many new SSR markers have been exploited constantly. A high-density ge-netic map was completed [6] based on the SSR markers in rice. Moreover, other molecular markers and a physical map have been constructed; especially the
XIA Jiu-Cheng et al.: Ultrastructure and Gene Mapping of the Albino Mutant al12 in Rice (Oryza sativa L.) 1113
draft sequence of the rice genome (O. sativa L. ssp. indica and O. sativa L. ssp. japonica) has been com-pleted[7,8]. These make it possible to clone the gene of the rice, resulting in the isolation of many important genes[9-13]. In this study, the gene al12 has been mapped and its biological mechanism has been dis-cussed.
1 Materials and Methods
1. 1 Materials
At the seedling stage, the albino phenomena was found in the offspring of a hybrid (D702B × DXiangB), and a stabilized albino mutant line (Line 259) was developed through seven consecutive gen-erations of selfing and selection. Both D702B and DxiangB are the indica maintenance lines. Line 9311 was used as a control because it is a model indica restorer line and has similar genetic background with the albino mutant under consideration.
1. 2 Methods
1. 2. 1 Temperature for expressing albino
Rice seeds were sown in a box and put into an RXZ-280B intelligent climate-control chamber (Jing- nan, Ningbo, China). The color changes of the rice seedlings were observed under different temperatures.
1. 2. 2 Time for expressing albino
Rice seeds were sown in the field at a temperature ranging between 18℃ and 24 and the color
℃
changes of the seedlings were observed at different stages after sowing.
1. 2. 3 Chlorophyll content
The chlorophyll content of the rice seedling was measured using MINOLTA (chlorophyll meter) SPAD-502 (MINOLTA, Osaka, Japan)[14].
1. 2. 4 Ultrastructure of the chloroplast
The leaves of the rice seedling were cut into small blocks (1 mm × 2 mm) and double fixation with 3% glutaraldehyde containing 0.1 mol /L phosphonic acid buffer (pH 7.3) and 1% osmic acid. These materials were dehydrated using an ethanol serials and were then embedded with epon812. Specimens were sliced using a microtome LKB-V (LKB, Uppsala, Sweden) and the chloroplast structure of the young leaf was observed using a transmission electron microscope H-600 (Hitachi, Hitachi, Japan)[15].
1. 2. 5 DNA extraction
DNA was extracted using the CTAB method[16].
1. 2. 6 SSR primer synthesis and PCR amplification
SSR primers were obtained from http://www. https://www.360docs.net/doc/9112069767.html,/microsat/ssr.html; and PCR amplifica-tion was performed as described previously[17].
1. 2. 7 Mapping
Individual plants in the mapping population (9311×259) were genotyped and the genetic maps were drawn using the Mapmaker3.0 software (Whitehead Institute for Biomedical Research Center, Massachusetts, USA).
1. 2. 8 RT-PCR
RNA was extracted from the equiponderant leaves of the mutant and 9311 using BIOZOL (Bio-Flux, Tokyo, Japan) and was then transcribed using Rever-TraAce (ToYoBo, Osaka, Japan); cDNA was amplified as follows: 94 for 5 min; 94 for
℃℃
1 min, 55 for 1 min, and 7
2 for 1 min, 30 cycles;
℃℃
and 72 for 6
℃ min. The PCR products were checked in 1.5% agaroses and each PCR amplification was repeated thrice.
2 Results
2. 1 Phenotyping of the albino mutant al12
The leaves of the albino mutant al12 were white before the 3-leaf-stage (less than 30 d after sowing) at a low temperature (<24℃) and then gradually turned green. The control line 9311 also exhibited a white color in the climate-controlled growth chamber when the temperature was lower than 16℃ (Tables 1 and 2, Figs. 1 and 2). The other normal varieties were also white at low temperatures.
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遗传学报 Acta Genetica Sinica V ol.33 No.12 2006
Table 1 Color change of young leaves of rice under different temperatures
Temperature( )
℃14 16 18 20 22 24 26 Albino mutant White White White White White White Green
9311 (CK) White White Green Green Green Green Green
Table 2 Color change of young leaves of rice after sowing seeds
Days after sowing (d) 10 20 30 40 Albino mutant White White Yellow Green
9311(CK) Green Green Green Green
Fig. 1 Phenotype of the albino mutant at the 2-leaf stage
Fig. 2 Phenotype of the 9311 at the 2-leaf stage
2.2 Chlorophyll content of the albino mutant
al12
The chlorophyll contents of the base, middle, and top leaves of the albino mutant grown in the cli-mate-control chamber were measured. When the mu-tant returned to normal, the chlorophyll content of the mutant and 9311, the check entry, were determined. The chlorophyll content of the mutant seedlings were significantly different from that of 9311, the check entry before the 3-leaf-stage (P< 0.01). There was no notable difference in the chlorophyll content between the mutant and 9311 (P < 0.01) when the mutant turned green, indicating that the chlorophyll content of the mutant had reached a normal level and that the mutant had also turned into normal (Table 3).
2. 3 Ultrastructure of the chloroplast of the mu-
tant al12
The ultrastructure of the mutant was observed at different temperatures (albino phenotype at 24℃ and green phenotype at 26℃) on a 2-leaf-stage using a transmission electron microscope. According to Figs. 3, 4, and 5, the chloroplast of the mutant did not develop normally, except for the formation of several small vesicle-like structures at 24℃. However, the mutant exhibited a normal chloroplast structure at 26℃ in the 2-leaf-stage. The observation showed that the ultrastructure of the mutant would become normal at 26℃.
XIA Jiu-Cheng et al .: Ultrastructure and Gene Mapping of the Albino Mutant al12 in Rice (Oryza sativa L.)
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Table 3 Chlorophyll content (CC) of the mutant and 9311
Seedling (2-leaf-stage)
Normal stage
Material Replication CC (SPAD ) Average (SPAD )
Replication
CC (SPAD ) Average (SPAD )
Mutant (259) 1 1.2 1 24.3 2 1.2 1.2 2 25.1 23.6 3 1.3 3 21.3 1 24.3 1 24.7 9311 (CK) 2 25.6 25.3 2 25.0 25.3 3 26.0 3 26.3 ANOV A. F (0.01)
2402.2**
1.2
**: significantly different level.
Fig. 3 Chloroplast of the mutant 259 at 24℃ (2-leaf-stage, × 35 000)
Fig. 4 Chloroplast of the mutant 259 at 24℃ (2-leaf-stage, × 15 000)
2.
4 Inheritance analysis of the albino mutant
al12 The albino mutant (259) was crossed and back-crossed with 9311 to bring about F 1 at Wenjiang, Chengdu in August, 2004. F 2 generation was obtained by self-crossing at Lingshui, Hainan. F 1 behaved green and F 2 fitted to the ratio 3:1 (green:white), as determined by the χ2-test (Table 4). This proved that the mutant was controlled by a recessive gene.
2. 5 Gene mapping of the albino mutant al12
A total of 44 polymorphic SSR markers between the mutant parent (259) and the parent 9311 were obtained from 350 pairs of the SSR primer (RM1- RM350). Using the bulked segregant analysis (BSA) method [18], it was observed that the SSR markers RM25, RM72, RM152, and RM339 had a linkage to the gene al12 besides the difference between the two DNA pools. In the mapping population (9311×259),
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Fig. 5 Chloroplast of the mutant 259 at 26℃(2-leaf-stage, ×35 000)
Table 4 Separation ratio among the F 2 population
Cross
Dominant
individual
Recessive individual
Total (3:1)
259×9311 594 189 783 0.269311×259 2116 719 2835 0.17
RM25 had 16 single-crossing recombinants and 1 double-crossing recombinant; RM72 had 26 sin-gle-crossing recombinants and 1 double-crossing re-combinant; RM152 had 30 single-crossing recombi-nants; and RM339 had 34 single-crossing recombi-nants and 1 double-crossing recombinant. The link-age analysis showed that the gene al12 was located on chromosome 8. RM152 and the other three mark-ers were located on the two flanks of the gene al12, respectively. The gene was separated from RM25, RM72, RM152, and RM339 by 12.3 cM, 26.0 cM, 18.4 cM, and 32.0 cM, respectively. Then 33 pairs of the SSR primer between RM25 and RM152 were synthesized [6]. Among these, the primers RM3702, RM5068, and RM8266 showed difference between
the two parents. RM3702 is a zero allele marker, which can be amplified as a clear band in 9311 but nothing in the mutant (259)[10]. To map exactly, the mapping population was extended to 300 recessive individuals. The primer RM3702 then had 24 sin-gle-crossing recombinants, RM5068 had 3 recombi-nants (zero allele markers cannot tell single-crossing or double-crossing), and RM8266 had 13 sin-gle-crossing recombinants and 1 double-crossing re-combinant. RM3702 and the other two markers, RM5068 and RM8266, stood on two flanks, sepa-rated from the gene by 4.9 cM, 0.5-1.1 cM, and 3.3 cM, respectively (Fig. 6). 2. 6 RT-PCR
According to the result of the mapping gene and the genome sequence of chromosome 8 (http://rgp. dna.affrc.go.jp), two candidate coding sequences (CDS) were found in the entire contigs between the two close SSR markers RM5068 and RM3702 on
chromosome 8 related with the mutant phenotype
Fig. 6 The part linkage map of the al12 gene on chromosome 8 of rice
XIA Jiu-Cheng et al.: Ultrastructure and Gene Mapping of the Albino Mutant al12 in Rice (Oryza sativa L.) 1117
(albino at low temperature): the putative cytochro-
mosome P450 (CYP) and the putative cold shock
protein (CSP). Three pairs of primers were designed
according to the sequence of the two CDSs. The in-
ternal reference was designed from β-actin (Table 5).
The RNA of the albino mutant and 9311 were retro-
transcripled into cDNA, and the cDNA were then
amplified using PCR. The result showed that the CDS
of exhibiting expression difference in the RNA level
is the putative cytochromosome P450 (CYP) (Fig. 7).
Table 5 Nucleic acid sequence of the four pairs of primers
Primers Forward (5′→3′) Reverse
(5′→3′)
CYP-1
CATTC CCAACA
CYP-2 ATTGCATGAGGCG-
AAGG
GCCCAGCAGCC-
AGAACA
CSP CACGCACTTATGA-
TGACC
AGTATGGACTAC-
GGAATGT
β-actin GAACTGGTATGG-
TCAAGGCTG
ACACGGAGCTCG-
TTGTAGAAG
Fig. 7 The result of RT-PCR of the mutant and 9311 with the four pairs of primers
Lane M: Marker; Lane 1, 3, 5, 7: mutant (259); Lane 2, 4, 6, 8: 9311.
3 Discussion
Albino of seedling resistant to low temperature, which is an important trait, has high value in biology and breeding. The energy for the development and growth of the young seedlings is provided by the en-dosperm before the 3-leaf-stage and thereafter by the solar energy. In young leaves of rice, at low tempera-ture, the chlorophyll synthesis and the development of chloroplast may temporarily stop to combat the adverse environment. After the 3-leaf-stage, the mu-tant resumes to synthesize chlorophyll and construct the chloroplast and thus begins to synthesize energy via photosynthesis. The albino mutant is a positive mutation. Except for the mutant al11 that behaves like a variegated albino and a greenable albino mutant W25, the other mutants reported are lethal mutants[1,2,5]. The new albino mutant al12 is similar to W25. Both are conditional expressions associated with the temperature and development stage[4] and are controlled by a recessive gene according to the separation behavior of F1 progeny plants[19]. However, these are different in their origins and preconditions of the albino expression between the line 259 and the line W25. The mutant W25 resulted from artificial mu- tation through irradiation of the thermosensitive male sterile line 2177s seeds with 300 Gy60Co-γray[4]. This mutant behaves like an albino before the 4-leaf-stage under 25℃. However, the mutant al12 originated from a natural mutation in the progenies of the hybrid D702B× DXiangB. Hence, it was consid- ered that the greenable albino mutant can be obtained by artificial and natural methods. Researches on the phenotype, physiology, biochemistry, and micro- structure have been carried out in the mutant W25[4,19,20]. In this study, the chlorophyll content of the albino mutant al12 was determined and the chloro- plast ultrastructure was observed. It was noted that the chlorophyll content was very low and the chloroplast was abnormal with only many micro-vesicles. Weng et al.[20] believed that the low content of soluble pro- tein and Rubisco, the lazy Rubisco activase, may be responsible for the albino.
In addition, the gene al12 was first located be-tween the two close SSR markers RM5068 and RM3702 on chromosome 8 by positional clon- ing[4,19,20]. Using RT-PCR, a CDS sequence of the putative cytochromosome P450 (CYP) exhibited ex-pression differences in the RNA level between the mutant and 9311. Cytochromosome P450 is a su-per-family including 10 subfamilies, such as CYP51, CYP71,CYP72, and CYP85, distributed in the whole genome of different plants[21]. This may be the key to the question regarding the existence of many different albino mutants in rice. Using Blast, it was observed that this putative sequence is very similar to the known gene CYP89A2 of Arabidopsis (http://www.
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遗传学报 Acta Genetica Sinica V ol.33 No.12 2006
https://www.360docs.net/doc/9112069767.html,/BLAST/), belonging to the CYP71 subfamily. The gene may associate with iron ion binding, oxygen binding, electron transport, amino acid and derivative metabolism, and lipid-metabolism (https://www.360docs.net/doc/9112069767.html,/entrez/query.fcgi?d b=gene&cmd=search&term=CYP89A2), (http:// https://www.360docs.net/doc/9112069767.html,/tigr-scripts/euk_manatee/shared/OR F_infopage.cgi?db=osa1&orf=11674.t00462). On the basis of this information, the mutant behaved al-bino at low temperatures in the seedling stage be-cause of the change of some base sequences in the open reading frame (ORF) or in the up/down stream of the CDS. However, further researche is necessary for fine mapping, cloning, transferring, and comple-mentation of the gene to confirm the necessity of al-bino mutation. Moreover, the albino gene al12 can be transferred into other varieties by continuous back-crossing, and new varieties with the albino marker can be bred to remove fake plants at early stages, to purify the seed, and to reduce the field work[22].
References:
[1] Iwata N, Satoh H, Omura T. Linkage analysis by use of
trisomics in rice(Oryza sativa L.). IV. Linkage groups
locating on chromosomes 2 and 10. Japan J Breed, 1981,
31 (Suppl. 1): 66-67.
[2] Iwata N, Omura T. Linkage studies in rice (Oryza satival)
some albino genes and their linkage relation with marker
genes. Sci Bull Fac Agr Kyus Hu Univ, 1978, 33(1): 18. [3] Maekawa K, Rikiishi K, Matsuura T, Noda K. Rever-
tants originated from chlorophyll mutants from the dis-
tantly related rice varieties. Japan J Breed Sci, 1996,
46(Suppl. 2): 107.
[4] SHU Qing-Yao, WU Dian-Xing, XIA Ying-Wu, LIU
Gui-Fu. Study on greenism characteristics of greenable
albino mutation line W25of rice (Oryza sativa L.).
Journal of Zhejiang Agricultural University, 1996, 22(2):
219-220 (in Chinese).
[5] YU Qing-Bo, JIANG Hua, MI Hua-Ling, ZHOU
Gen-Yu, YANG Zhong-Nan. The physiological character
and molecular mapping in rice albino21 mutant. Journal
of Shanghai NormaUniversity, 2005, 34(1): 70-75(in
Chinese with an English abstract).
[6] McCouch S R, Teytelman L, Xu Y, Lobos K B, Clare K,
Walton M, Fu B, Maghirang R, Li Z, Xing Y Z, Zhang Q
F, Kono I, Yano M, Fjellstrom R, DeClerck G, Schneider
D, Cartinhour S, Ware D, Stein L. Development and
mapping of 2240 new SSR markers for rice (Oryza sa-
tiva L.). DNA Research, 2002, 9: 199-207.
[7] Yu J, Hu S, Wang J, Wong G K, Li S, Liu B, Deng Y,
Dai L, Zhou Y, Zhang X, Cao M, Liu J, Sun J, Tang J,
Chen Y, Huang X, Lin W, Ye C, Tong W, Cong L, Geng J,
Han Y, Li L, Li W, Hu G, Huang X, Li W, Li J, Liu Z, Li
L, Liu J, Qi Q, Liu J, Li L, Li T, Wang X, Lu H, Wu T,
Zhu M, Ni P, Han H, Dong W, Ren X, Feng X, Cui P, Li
X, Wang H, Xu X, Zhai W, Xu Z, Zhang J, He S, Zhang
J, Xu J, Zhang K, Zheng X, Dong J, Zeng W, Tao L, Ye J,
Tan J, Ren X, Chen X, He J, Liu D, Tian W, Tian C, Xia
H, Bao Q, Li G, Gao H, Cao T, Wang J, Zhao W, Li P,
Chen W, Wang X, Zhang Y, Hu J, Wang J, Liu S, Yang J,
Zhang G, Xiong Y, Li Z, Mao L, Zhou C, Zhu Z, Chen R,
Hao B, Zheng W, Chen S, Guo W, Li G, Liu S, Tao M,
Wang J, Zhu L, Yuan L, Yang H. A draft sequence of the
rice genome (Oryza sativa L. ssp. indica). Science, 2002,
296(5565): 79-92.
[8] Goff S A, Ricke D, Lan T H, Presting G, Wang R, Dunn
M, Glazebrook J, Sessions A, Oeller P, Varma H, Hadley
D, Hutchison D, Martin C, Katagiri F, Lange B M,
Moughamer T, Xia Y, Budworth P, Zhong J, Miguel T,
Paszkowski U, Zhang S, Colbert M, Sun W L, Chen L,
Cooper B, Park S, Wood T C, Mao L, Quail P, Wing R,
Dean R, Yu Y, Zharkikh A, Shen R, Sahasrabudhe S,
Thomas A, Cannings R, Gutin A, Pruss D, Reid J, Tav-
tigian S, Mitchell J, Eldredge G, Scholl T, Miller R M,
Bhatnagar S, Adey N, Rubano T, Tusneem N, Robinson
R, Feldhaus J, Macalma T, Oliphant A, Briggs S. A draft
sequence of the rice genome (Oryza sativa L. ssp. in-
dica). Science, 2002, 296(5565): 92-100.
[9] LI Xue-Yong, QIAN Qian, FU Zhi-Ming, WANG
Yong-Hong, XIONG Guo-Sheng, ZENG Da-Li, WANG
Xiao-Qun, LIU Xin-Fang, TENG Sheng, Hiroshi Fuji-
moto, YUAN Ming, LUO Da, HAN Bin, LI Jia-Yang.
Control of tillering in rice. Nature, 2003. 422(6932):
618-621.
[10] LI Pei-Jin, ZENG Da-li, LIU Xin-Fang, XU Dan, GU
Dai, LI Jia-Yang, QIAN Qian. Heredity and mapping of
a lazy growth mutant in rice (Oryza sativa L.). Chinese
Science Bulletin, 2003, 48(21): 2271-2274 (in Chinese).
[11] WANG Yun, XIAO Han, QIAN Qian, LI Hong-Chang,
LI Shi-Gui, ZHU Li-Huang. Genetical studies and fine
mapping of lax panicles mutant in rice (Oryza sativa L.).
Chinese Science Bulletin, 2003, 48(15): 1666-1670 ( in
Chinese ).
[12] LIANG Guo-Hua, CAO Xiao-Ying, SUI Jiong-Ming,
ZHAO Xiang-Qiang, YI Chuan-Deng, GU Ming-Hong.
Fine mapping of a semidwarf gene sd-g in rice. Chinese
Science Bulletin, 2004, 49(8): 778-783 ( in Chinese ). [13] WANG Ren-Xiao, LI Pei-Jin, CHEN Hong-Qi, WEN
Shao-Kai, LI Jia-Yang, ZHU Xu-Dong. Fine Localiza-
XIA Jiu-Cheng et al.: Ultrastructure and Gene Mapping of the Albino Mutant al12 in Rice (Oryza sativa L.) 1119
tion of rice EUI1 gene controlling elongation of the up-
permost intermode. Acta Genetica Sinica, 2005, 32(9): 955-959 ( in Chinese with an English abstract).
[14] MA Jun, ZHU Qing-Sen, MA Wen-Bo, TIAN Yan-Hua,
YANG Jian-Chang, ZHOU Kai-Da. Studies on the pho-
tosynthetic characteristics and accumulation and trans-
formation of assimilation product in heavy panicle type of rice. Scientia Agricultura Sinica, 2003, 36(4): 375- 381 (in Chinese with an English abstract).
[15] SHAO Ji-Rong, WANG Yu-Zhong, LIU Yong-Sheng,
SUN Jing-San, XIE Rong. Ultrastructural observation on
chloroplast of the thermo-sensitive-green-yellow banded
leaf mutant in rice (Oryza sativa L. ssp. indica). Acta
Botanica Sinica, 1999, 41 (1): 20-24 (in Chinese with an
English abstract).
[16] Murray M G, Thompson W F. Rapid isolation of high
molecular weight plant DNA. Nucleic Acids Res, 1980, 8(19): 4321-4325.
[17] Panaud O, Chen X, MoCouch S R. Development of mi-
crosatellite makers and characterization of simple se-
quence length polymorphisms(SSLPs) in rice (Oryza sa-
tiva L.). Mol Gen Genet, 1996, 252(5): 597-607. [18] Michelmore R W, Paranand I, Kessali R V. Identification
of markers linked to disease resistance gene by bulked
segregant analysis: a rapid method to detect markers in specific genomic regions using segregating populations.
Proc Natl Acad Sci USA, 88: 9829-9832.
[19] WU Dian-Xing, SHU Qing-Yao, XIA Ying-Wu, LIU
Gui-Fu, LI Jun-Ying. Study on Chloroplast ultrastructure
of a greenable albine mutation line cv. W25 of rice (Oryza sativa). Journal of Zhejiang Agricultural Univer-
sity, 1997, 23 (4): 451-452 (in Chinese).
[20] WENG Xiao-Yan, JIANG De-An, LU Qing. Changes in
rubisco activity, Rubisco activase activity and photosyn-
thetics rate in greenable albino mutation line of rice dur-
ing greening. Acta Phytophysiologica Sinica, 2000, 26(3): 213-218 (in Chinese with an English abstract). [21] Nelson D R, Schuler M A, Paquette S M, Werck-Reich-
hart D, Bak S. Comparative genomics of rice and arabi-
dopsis. analysis of 727 cytochrome P450 genes and pseudogenes from a monocot and a dicot. Plant Physiol,
2004, 135: 756―772.
[22] Cao Li-Yong, Qian Qian, Zhu Xu-Dong, Zeng Da-Li ,
Min Shao-Kai, Xiong Zhen-Min. Breeding of a photo-
thermo sensitive genic male sterile indica rice ZhongziS
with a purple-leaf marker and the heterosis of its hybrid
rice produced with it . Acta Agronomica Sinica, 1999, 25
(1): 39-43 (in Chinese with an English abstract).
水稻(Oryza sativa L.)苗期低温白化突变体al12的超微结构
与基因定位
夏九成1,2,3，王玉平1,2，马炳田1,2，殷兆晴1,2，郝铭1,2，孔德伟1,2，李仕贵1,2
1. 四川农业大学水稻研究所，温江 611130；
2. 作物基因资源与遗传改良教育部重点实验室，雅安 625014；
3. 四川省攀枝花学院, 攀枝花 617000
摘要:水稻苗期低温白化突变是水稻在发育早期对低温胁迫的一种适应性，是一种受发育和温度控制的条件表达，它与其他水稻白化突变有本质的不同。本研究利用便携式叶绿素测量仪测定了白化时期植株的叶绿素含量和用透射电镜观察了叶绿体的结构变化。结果发现叶绿素平均含量仅为1.2（SPAD），而叶绿体也不能正常发育仅有囊泡状结构。通过与9311的正反交实验及子代的分离表现证明该性状受一个隐性核基因的控制。另外利用SSR分子标记技术将该基因定位在第8染色体上，两侧最近的SSR标记RM5068和RM3702分别距基因0.5～1.1 cM和4.9 cM，基因被定位在约6个cM的区间内。我们将该基因暂时命名为al12。
关键词:水稻；白化突变体；叶绿体；基因定位
作者简介: 夏九成（1976-），男，四川西昌人，在读博士，研究方向：植物遗传学。E-mail: xjch04@https://www.360docs.net/doc/9112069767.html,
水稻(Oryza sativa L.)苗期低温白化突变体al12的超微结构与基因定位
作者：夏九成， 王玉平， 马炳田， 殷兆晴， 郝铭， 孔德伟， 李仕贵， XIA Jiu-Cheng， WANG Yu-Ping， MA Bing-Tian， YIN Zhao-Qing， HAO Ming， KONG De-Wei， LI Shi-Gui
作者单位：夏九成,XIA Jiu-Cheng(四川农业大学水稻研究所,温江,611130;作物基因资源与遗传改良教育部重点实验室,雅安,625014;四川省攀枝花学院,攀枝花,617000)， 王玉平,马炳田,殷兆晴,郝铭,孔德伟,李仕贵,WANG Yu-
Ping,MA Bing-Tian,YIN Zhao-Qing,HAO Ming,KONG De-Wei,LI Shi-Gui(四川农业大学水稻研究所,温江
,611130;作物基因资源与遗传改良教育部重点实验室,雅安,625014)
刊名：
遗传学报
英文刊名：ACTA GENETICA SINICA
年，卷(期)：2006,33(12)
被引用次数：10次
参考文献(22条)
1.Iwata N;Satoh H;Omura T Linkage analysis by use of trisomics in rice(Oryza sativa L.).Ⅳ.Linkage groups locating on chromosomes 2 and 10 1981(z1)
2.Iwata N;Omura T Linkage studiesin rice (Oryza satival)some albino genes and their linkage relation with marker genes 1978(01)
3.Maekawa K;Rikiishi K;Matsuura T;Noda K Revertants originated from chlorophyll mutants from the distautly related rice varieties 1996(z2)
4.SHU Qing-Yao;WU Dian-Xing;XIA Ying-Wu;LIU Gui-Fu Study on greenism characteristics of greenable albino mutation line W25 of rice (Oryza sativa L.) 1996(02)
5.YU Qing-Bo;JIANG Hua;MI Hua-Ling;ZHOU Gen-Yu YANG Zhong-Nan The physiological character and molecular mapping in rice albino21 mutant[期刊论文]-Journal of Shanghai NormaUniversity 2005(01)
6.McCouch S R;Teytelman L;Xu Y;Lobos K B,Clare K,Walton M,Fu B,Maghirang R,Li Z,Xing Y Z,Zhang Q F,Kono I,Yano
M,Fjellstrom R,DeClerck G,Schneider D,Cartinhour S,Ware D,Stein L Development and mapping of 2240 new SSR markers for rice (Oryza sativa L.)[外文期刊] 2002
7.Yu J;Hu S;Wang J;Wong G K,Li S,Liu B,Deng Y,Dai L,Zhou Y,Zhang X,Cao M,Liu J,Sun J,Tang J,Chen Y,Huang X,Lin W,Ye C,Tong W,Cong L,Geng J,Hah Y,Li L,Li W,Hu G,Huang X,Li W,Li J,Liu Z,Li L,Liu J,Qi Q,Liu J,Li L,Li T,Wang X,Lu H,Wu
T,Zhu M A draft sequence of the rice genome (Oryza sativa L.ssp.indica)[外文期刊] 2002(5565)
8.Goff S A;Ricke D;Lan T H;Presting G,Wang R,Dunn M,Glazebrook J,Sessions A,Oeller P,Varma H,Hadley D,Hutchison
D,Martin C,.Katagiri F,Lange B M,Moughamer T,Xia Y,Budworth P,Zhong J,Miguel T,Paszkowski U,Zhang S,Colbert M,Sun W
L,Chen L,Cooper B,Park S,Woo A draft sequence of the rice genome (Oryza sativa L.ssp.indica)[外文期刊] 2002(5565)
9.LI Xue-Yong;QIAN Qian;FU Zhi-Ming;WANG Yong-Hong,XIONG Guo-Sheng,ZENG Da-Li,WANG Xiao-Qun,LIU Xin-Fang,TENG
Sheng,Hiroshi Fujimoto,YUAN Ming,LUO Da,HAN Bin,LI Jia-Yang Control of tillering in rice[外文期刊] 2003(6932)
10.LI Pei-Jin;ZENG Da-li;LIU Xin-Fang;XU Dan,GU Dai,LI Jia-Yang,QIAN Qian Heredity and mapping of a lazy growth mutant in rice (Oryza sativa L.)[期刊论文]-Chinese Science Bulletin 2003(21)
11.WANG Yun;XIAO Han;QIAN Qian;LI Hong-Chang,LI Shi-Gui,ZHU Li-Huang Genetical studies and fine mapping of lax panicles mutant in rice (Oryza sativa L.)[期刊论文]-Chinese Science Bulletin 2003(15)
12.LIANG Guo-Hua;CAO Xiao-Ying;SUI Jiong-Ming;ZHAO Xiang-Qiang YI Chuan-Deng GU Ming-Hong Fine mapping of a semidwarf gene sd-g in rice[期刊论文]-Chinese Science Bulletin 2004(08)
13.WANG Ren-Xiao;LI Pei-Jin;CHEN Hong-Qi;WEN Shao-Kai LI Jia-Yang ZHU Xu-Dong Fine Localization of rice EUI1 gene controlling elongation of the uppermost intermode[期刊论文]-Acta Genetica Sinica 2005(09)
14.MA Jun;ZHU Qing-Sen;MA Wen-Bo;TIAN Yan-Hua YANG Jian-Chang ZHOU Kai-Da Studies on the photosynthetic characteristics and accumulation and transformation of assimilation product in heavy panicle type of rice[期刊论文]-Scientia
17.Panaud O;Chen X;MoCouch S R Development of microsatellite makers and characterization of simple sequence length polymorphisms(SSLPs) in rice (Oryza sativa L.)[外文期刊] 1996(05)
18.Michelmore R W;Paranand I;Kessali R V Identification of markers linked to disease resistance gene by bulked segregant analysis:a rapid method to detect markers in specific genomic regions using segregating populations
19.WU Dian-Xing;SHU Qing-Yao;XIA Ying-Wu;LIU Gui-Fu,LI Jun-Ying Study on Chloroplast ultrastructure of a greenable albine mutation line cv.W25 of rice(Oryza sativa) 1997(04)
20.WENG Xiao-Yan;JIANG De-An;LU Qing Changes in rubisco activity,Rubisco activase activity and photosynthetics rate in greenable albino mutation line of rice during greening 2000(03)
21.Nelson D R;Schuler M A;Paquette S M;Werck-Reichhart D Bak S Comparative genomics of rice and arabidopsis,analysis of 727 cytochrome P450 genes and pseudogenes from a monocot and a dicot[外文期刊] 2004
22.Cao Li-Yong;Qian Qian;Zhu Xu-Dong;Zeng Da-Li Min Shao-Kai Xiong Zhen-Min Breeding of a photothermo sensitive genic male sterile indica rice ZhongziS with a purple-leaf marker and the heterosis of its hybrid rice produced with it
1999(01)
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3.吴军.陈佳颖.赵剑.林冬枝.董彦君2个水稻温敏感叶色突变体的光合特性研究[期刊论文]-中国农学通报 2012(21)
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2010(3)
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10.董华林.费震江.魏磊.武晓智.周鹏水稻苗期叶片白化转绿性状研究进展[期刊论文]-湖北农业科学 2012(23)
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