Download Expressed Sequence Tags from Citrus Albedo at the Initiation Stage

FUJII etSci.
al.:2Expressed
Sequence
Bull. Natl. Inst. Fruit Tree
: 127 ∼ 144,
2003 Tags from Citrus Albedo at the Initiation Stage of Rind Peeling
127
Research note
Expressed Sequence Tags from Citrus Albedo
at the Initiation Stage of Rind Peeling
Hiroshi FUJII, Masayuki KITA†1, Takehiko SHIMADA, Tomoko ENDO and Mitsuo OMURA
Department of Citrus Research, Okitsu, National Institute of Fruit Tree Science
National Agricultural Research Organization, Shimizu, Shizuoka 424-0292, Japan
Abstract
The gene expression and regulation associated with the initial stage of rind peeling of the satsuma
mandarin (Citrus unshiu cv. ‘Miyagawa-wase’) was characterized through the generation of 626 expressed
sequence tags (ESTs) derived from a cDNA library prepared from albedo tissues collected 60 days after
anthesis. A Total of 451 ESTs was found to have significant similarity to the known sequences. Using our inhouse database, a catalogue of 222 ESTs annotated with their putative functions was prepared and compared
to the previously prepared catalogue of 442 ESTs derived from mature albedo tissues. In consequence, gene
homologues associated with cell wall events were preferably found in this library.
Key words: Citrus unshiu; ESTs; cDNA library, gene catalogue
fruits, we have completed the EST analysis of several
Introduction
cDNA libraries derived from sweet orange young seeds
(Hisada et al., 1996), juice sac of satsuma mandarin during
The analysis of expressed sequence tags (ESTs) has
rapid cell development phase (Hisada et al., 1997), satsuma
become a part of the genomic approach to plant science
mandarin fruit at mature phase (Moriguchi et al., 1998),
for many plant species (Yamamoto and Sasaki, 1997).
albedo of satsuma mandarin at mature phase (Kita et al.,
The cataloguing of ESTs has emerged as a powerful tool
2000) and ovary at the day of anthesis (Shimada et al., in
capable of revealing gene expression patterns, gene
press). We are particularly interested in the traits that
regulation and sequence diversity (Brandle et al., 2002)
contribute to the quality and commercial value of citrus
as well as assisting in the generation of DNA markers for
fruits. It is considered that easy-peeling, which is one of
genome mapping (Kuhara et al., 1994). In most cases it
the important traits for commercial value in citrus, is
has also given an indispensable aid to cDNA microarray
related with the relaxation of cell wall in particular
analysis. Furthermore, EST databases have recently
(Brummell and Harpster, 2001). Therefore, we have
contributed to the discovery of the specific gene
catalogued EST derived from albedo of satsuma mandarin
repertories associated with the fragrances of roses
fruits at the initiation stage of peeling of the rind in which
(Guterman et al., 2002) and terpenoids in Stevia redaudiana
the genes related to cell wall relaxation should be highly
(Brandle et al., 2002).
expressed. In this report, we list 222 cDNA clones, which
In order to understand gene expression and regulation
were identified by their putative functions by database
associated with the development and ripening of citrus
searches and discuss the characteristic features of the
†1
The catalogued cDNA clones have been submitted to DDBJ
under accession numbers AU300309-AU300928.
Present address: Department of Plant Cell and Environment,
National Institute of Fruit Tree Science, Tsukuba, Ibaraki 3058605, Japan
Bull. Natl. Inst. Fruit Tree Sci. 2. 2003
128
catalogue.
wase’) were sequenced. The 626 ESTs generated form
the library were clustered into 541 independent groups.
Materials and Methods
The average insert size of the resulting ESTs was on ca.
440bps. Comparison of the sequences with ESTs in the
1. Construction of cDNA library
DDBJ and our in-house citrus EST database resulted in
‘Miyagawa-wase’ variety of satsuma mandarin (Citrus
the identification of 451 sequences with significant
unshiu Marc.), which was cultivated at the National
homology to known sequences. Among them, 336 ESTs
Institute of Fruit Tree Science, Okitsu (Shimizu, Shizuoka,
(53.7%) could be functionally identified, 90 ESTs (14.3%)
Japan), was used as all fruit sample in this study. Samples
had homology to EST from other organisms and 25 ESTs
were collected 60 days after anthesis (July 16, 1999)
(4.0%) shared homology with only putative open reading
when the albedo (inner spongy portion) was capable of
frame of Arabidopsis thaliana genomic sequences. There
being peeled from the segment membrane. The average
were only 10 ESTs with homology to genes of other
fruit size was 26.2 mm in diameter with a fresh weight of
citrus species (Table 1). Significant similarities were not
9.0 g. Albedo tissue was peeled from the segment
detected for the remaining 175 ESTs (28.0%). As a cause
membrane using a razor, immediately frozen with liquid
of no similarity, it was considered that either the sequences
nitrogen, and stored at – 80 ℃. The construction of the
of 3’ untranslated region were obtained, or ESTs were
cDNA library and the subsequent sequencing were
derived from unknown genes which had never registered
conducted following the method of Kita et al. (2000). The
in the DDBJ. The ALP EST data will be registered in the
cDNA library derived from pre-mature albedo tissue was
DDBJ for the public use. Two hundred and twenty two
designated as the ALP cDNA library.
ESTs were catalogued and the resulting ESTs are shown
in Table 1 with their putative functions and homology
2. Data processing and cataloguing
search results. Even in case when isozymes of the same
A total of 626 cDNA clones were randomly selected
function were found among the above 336 ESTs identified
and sequenced from both the 5’ and 3’ ends of the
the functions, a single example was entered into the
inserts. Each sequence was compared with the nucleic
catalogue.
acid sequences in the DDBJ (http://www.ddbj.nig.ac.jp/;
release 51, Sep. 2002) and also with our in-house citrus
2. Gene repertories of ALP cDNA library
EST database (Fujii et al., 2003) using the FASTA algorithm
ESTs in ALP cDNA library were classified according to
(Pearson and Lipman, 1988). A significant match was
putative function (Table 2). In this library, 20 gene
declared when the optimized similarity score (OPT) was
homologues associated with cell wall events were found.
greater than 400. ESTs information including sequence
Changes in the cell wall including expansion of the cell wall
data and annotations were entered into our in-house
may possibly be related to the easy-peeling characteristic of
citrus EST database. Following annotation, a catalogue of
the satsuma mandarin. Since morphological changes in
222 ESTs based on function was prepared. Preparation of
albedo would be related to the reduction of pectic
the cDNA library and catalogue derived from mature
substances (Kuraoka and Kikuchi, 1961), homologues of
albedo tissue designated as ALM was previously reported
pectate lyase (MAP9U30), pectin acetylesterase (MAPEM60),
(Kita et al., 2000). Data analysis, including the comparison
pectin methylesterase (MAPAE10), polygalacturonase
of the ALP and ALM catalogues was carried out using the
(MAP9U96), arabinogalactan protein (MAPAE91: Shultz
in-house database.
et al., 2000) were found and they might be related to such
an event. Homologue of glycine-rich protein (MAPAE88:
Results and Discussion
Uthappa et al., 2001) and proline-rich protein (MAPAE26:
Salts et al., 1992) are possibly related to cell structure
1. Sequence Analysis
and cell adhesion in the cell wall. Homologues associated
A total of 626 cDNA clones from the ALP cDNA library
to cellulose, which is a major component of cell wall
derived from albedo tissue of satsuma mandarin (‘Miyagawa-
structure, were also detected (MAPF194, MAPH163:
FUJII et al.: Expressed Sequence Tags from Citrus Albedo at the Initiation Stage of Rind Peeling
Favery et al., 2001). The above-mentioned MAPEM60 and
MAP9U96 had two redundant ESTs each in total of 626
129
Sci. 122: 808-812.
8）Kita, M., T. Endo, T. Moriguchi and M. Omura. 2000. cDNA
ESTs and were constructed contig sequences respectively.
catalogs expressed in albedo of citrus fruit: a comparative
When the ALP gene catalogue was compared to a
analysis of cDNA libraries from pulp and albedo of
catalogue ALM derived from mature albedo tissue, only
satsuma mandarin (Citrus unshiu Marc.). Acta Hort. 521:
44 kinds (80 clones) of 336 gene homologues in ALP were
179-183.
overlapped. Several examples of them were dynamin-like
9）Kuhara, N., Y. Nagamura, K. Yamamoto, Y. Harushima,
protein (MAPAE24), ubiquitin SMT3 protein (MAPAE81),
N. Sue, J. Wu, B.A. Antonio, A. Shormura, T. Shimizu, S-Y.
cystein proteinase (MAPAT76), actin (MAPDD12) and
Lin, T. Inoue, A. Fukuda, T. Shimano, Y. Kuboki, T.
thioredoxin (MAPFF53). Since the percentage of the
Toyama, Y. Miyamoto, T. Kirihara, K. Hayasaka, A. Miyano,
overlapped clones is low, the ALP library is suitable to for
L. Monna, H.S. Zhong, Y. Tamura, Z-X. Wang, T. Monna, Y.
the exploration of important genes associated with the
Umehara, M. Yano, T. Sasaki and Y. Minobe. 1994. A 300
maturation of albedo.
kilobase interval genetic map of rice including 883
expressed sequences. Nature Genet. 8: 365-372.
Literature Cited
1）Brandle, J. E., A. Richman, A. K. Swanson and B. P. Chapman.
2002. Leaf ESTs from Stevia redaudiana: a resource for
gene discovery in diterpene synthesis. Plant Mol. Biol.
50: 613-622.
10）Kuraoka, T. and T. Kikuchi. 1961. Morphological studies
on the development of citrus fruits. I. Satsuma orange. J.
Japan. Soc. Hort. Sci. 30: 189-196.
11）Moriguchi, T., M. Kita, S. Hisada, T. Endo and M. Omura.
1998. Characterization of gene repertories at mature stage
2）Brummell, D. A. and M. H. Harpster. 2001. Cell wall
of citrus fruits through random sequencing and analysis
metabolism in fruit softening and quality and its manipulation
of redundant metallothionein-like genes expressed during
in transgenic plants. Plant Mol. Biol. 47: 311-340.
fruit development. Gene 211: 221-227.
3）Favery, B., E. Ryan, J. Foreman, P. Linstead, K. Boudonck,
12）Pearson, W.R. and D.J. Lipman. 1988. Improved tools for
M. Steer, P. Shaw and L. Dolan. 2001. KOJAK encodes a
biological sequence comparison. Proc. Natl. Acad. Sci.
cellulose synthase-like protein required for root hair cell
morphogenesis in Arabidopsis. Genes Dev. 15: 79-89.
USA 85: 2444-2448.
13）Salts, Y., R. Wachs, D. Kenigsbuch, W. Gruissem and R.
4）Fujii, H., T. Shimada, T. Endo and M.Omura. 2003.
Barg. 1992. DNA sequence of the tomato fruit expressed
Development of relational database system for citrus ESTs.
proline-rich protein gene TPRP-F1 reveals an intron within
Bull. Natl. Inst. Fruit Tree Sci. 2: 91-99.
the 3 untranslated transcripts. Plant Mol. Biol. 18: 407-409.
5）Guterman, I., M. Shalit, N. Menda, D. Piestun, M. Dafny-Yelin,
14）Schultz, C.J., K.L. Johnson, G. Currie and A. Bacic. 2000.
G. Shalev, E. Bar, O. Davydov, M. Ovadis, M. Emanuel, J.
The classical arabinogalactan protein gene family of
Wang, Z. Adam, E. Pichersky, E. Lewinsohn, D. Zamir, A.
Arabidopsis. Plant Cell 12: 1751-1767.
Vainstein and D. Weiss. 2002. Rose scent: Genomics
15）Shimada, T., M. Kita, T. Endo, H. Fujii, T. Ueda, T.
approach to discovering novel floral fragrance-related genes.
Moriguchi and M. Omura. Expressed Sequence Tags of
Plant Cell 14: 2325-2338.
ovary tissue cDNA library in Citrus unshiu Marc. Plant
6）Hisada, S., T. Moriguchi, T. Hidaka, A. M. Koltunow, T.
Science in press.
Akihama and M. Omura. 1996. Random sequencing of sweet
16）Uthappa, M., U.S. Muchhal, J.C. Baldwin and K.G.
orange (Citrus sinensis Osbeck) cDNA library derived
Raghothama. 2001. LeGRP1: A new member of glycine-rich
form young seeds. J. Japan. Soc. Hort. Sci. 65: 487-495.
proteins from tomato (Lycopersicon esculentum). Physiol.
7）Hisada, S., T. Akihama, T. Endo, T. Moriguchi and M.
Omura. 1997. Expressed sequence tags of citrus fruit
during rapid cell development phase. J. Amer. Soc. Hort.
Plant. 113: 85-91.
17）Yamamoto, K. and T. Sasaki. 1997. Large-scale EST
sequencing in rice. Plant Mol. Biol. 35: 135-144.
21kD cell wall protein
2C-methyl-D-erythritol 2,4-cyclodiphosphate synthase (MECS)
3-deoxy-D-manno-octulosonic acid transferase-like protein
(MED24.5)
4-methyl-5(b-hydroxyethyl)-thiazole monophosphate biosynthesis
protein putative
MAPH109 AU300763
MAPEM44 AU300848
MAPDD68 AU300899
MAPF102
actin depolymerizing factor
adenine nucleotide translocator 1 (CANT1)
adenylosuccinate synthase
ADP-ribosylation factor
aldolase (SCA1) cytosolic
MAPGL96 AU300718
MAPDD32 AU300839
MAPBG79 AU300812
MAPDR28 AU300473
MAPFF16
aquaporin PIP1-3 (PIP1-3) putative
arabinogalactan protein (AGP16)
arbutin synthase
MAPBG63 AU300421
MAPAE91 AU300917
MAP9U70
MAPGL32 AU300927
aspartic protease precursor
aquaporin PIP1-1 (PIP1-1) putative
MAPH120 AU300765
AU300348
α-cpn60 precursor (alphacpn60)
MAPDD41 AU300914
AU300632
actin
MAPDD12 AU300432
AU300900
14-3-3 protein
MAPEF48 AU300517
Putative identity / Homolog
13-lipoxygenase
Accession
MAPGL13 AU300674
Clone
stress
vitamin
lipid/membrane
formation
membrane protein
membrane protein
folding
sugar
trafficking/G-protein
nucleic acid synthesis
Protein
regulation
Metabolism
proteinase
secondary
Cell structure cell wall
Membrane
Membrane
Protein
regulation
Metabolism
Protein
regulation
Metabolism
nucleic acid
Cell structure skelton
Metabolism
second
secondary
Cell structure skelton
Metabolism
Metabolism
Metabolism
Cell structure cell wall
Unclassified
Responsive
first
Category of Homolog
L46681
AJ310148
AF195897
AF141899
AF141643
U21105
AF308587
M95166
U49389
AF006489
AF183904
X68649
AF326856
AY065115
AF250236
Y11553
AF039709
X96405
Accession of
Homolog
705
1234
704
613
500
1469
711
453
846
1345
571
1353
1057
opt
Lycopersicon
esculentum
Rauvolfia serpentina
Arabidopsis thaliana
647
567
479
Vitis berlandieri x Vitis 1142
rupestris
Vitis berlandieri x Vitis 1230
rupestris
Pisum sativum
Fragaria x ananassa
Arabidopsis thaliana
Arabidopsis thaliana
Gossypium hirsutum
Petunia x hybrida
Pisum sativum
Arabidopsis thaliana
Arabidopsis thaliana
Catharanthus roseus
Medicago sativa
Maackia amurensis
Solanum melongena
Organism
Table 1. Inventory of C. unshiu ESTs with significant similarity to deduced genes from other organisms
66.5
69.8
64.7
81.0
78.9
82.8
81.5
87.3
74.0
71.2
81.9
84.1
72.3
67.8
66.9
66.0
81.0
58.4
Identity
(%)
337
255
286
352
408
204
368
181
227
208
442
201
177
423
755
353
411
1013
Over
lap
130
Bull. Natl. Inst. Fruit Tree Sci. 2. 2003
β-tubulin 4
β-tubulin 5 (Tubb5)
MAPAT93 AU300403
MAPFF37
Ca2+/H+ exchanger
calcium binding protein
MAPGR85 AU300846
MAP9U41
carboxypeptidase II, serine-type putative
catalase
MAPEF07 AU300501
AU300635
AU300856
MAPFF21
MAP9U22
cellulose synthase catalytic subunit (celA3)
MAPF194
centrin
MAPFF60
chitinase-like protein 1 (CTL1)
AU300537
AU300853
MAPEI58
MAPEI91
chlorophyll a/b-binding protein
chaperonin 60
MAPGL63 AU300845
AU300842
cellulose synthase like CSLD3 (CslD3) involved in biosynthesis of
non-cellulosic beta-glucan-containing polysacchride
MAPH163 AU300811
AU300624
CBL-interacting protein kinase 3 (CIPK3)
MAPH180 AU300820
catechol O-methyl transferase 5
calmodulin
MAPGR59 AU300743
AU300338
biotin carboxyl carrier protein subunit precursor(accB-2), of acetyl
CoA carboxylase
MAPEF36 AU300512
AU300642
β-hydroxybutyryl-CoA dehydrogenase(hbdA)
MAPGL79 AU300710
Putative identity / Homolog
β-1,3-glucanase putative
Accession
MAPAE07 AU300880
Clone
Table 1. Continued
lipid
stress
proteinase
calcium
calcium
calcium
Metabolism
Responsive
Protein
regulation
Responsive
photosystem
defense
folding
defense/cytoskelton
Cell structure cell wall
Cell structure cell wall
Signal
second
transport
lipid
Cell structure lignin
Responsive
Protein
regulation
Signal
Signal
Membrane
Metabolism
Cell structure skelton
Cell structure skelton
Metabolism
Cell structure cell wall
first
Category of Homolog
M85150
AF422179
X70867
AJ009672
AF232907
AF150630
AF286051
AF064695
AF151368
AF370244
X60738
AJ309072
AB012932
AF271071
U76896
D63136
U32229
AY065273
Accession of
Homolog
Arabidopsis thaliana
Arabidopsis thaliana
Cucurbita sp.
Arabidopsis thaliana
Arabidopsis thaliana
Gossypium hirsutum
Arabidopsis thaliana
Thalictrum tuberosum
Hevea brasiliensis
Arabidopsis thaliana
Malus domestica
Sesbania rostrata
Vigna radiata
Glycine max
Triticum aestivum
Zinnia elegans
Bradyrhizobium
japonicum
Arabidopsis thaliana
Organism
778
706
1403
1235
869
1564
1210
445
703
570
732
425
918
607
1734
806
520
856
opt
75.5
74.0
80.9
77.8
73.1
88.1
75.2
61.9
74.2
72.0
84.3
70.9
70.9
79.2
85.4
71.3
62.4
71.8
Identity
(%)
278
281
430
427
334
395
431
320
275
225
204
172
381
197
471
373
338
341
Over
lap
FUJII et al.: Expressed Sequence Tags from Citrus Albedo at the Initiation Stage of Rind Peeling
131
cinnamate 4-hydroxylase
cinnamyl alcohol dehydrogenase
MAPEM35 AU300557
AU300769
AU300547
AU300818
MAPH142
MAPEI93
MAP9U91
coatomer epsilon subunit protein putative
copper chaperone homolog
cp protein IM30 protein
cystatin-like protein
cysteine proteinase putative
MAPAE52 AU300380
MAPEM06 AU300815
MAPGR94 AU300823
MAPEM59 AU300569
AU300915
AU300404
MAPAT76
MAPAT95
cytochrome P-450, n-alkane inducible ALK6-A (CYP52C2)
MAP9U84
dihydroflavonol reductase
MAPEF53
AU300518
dehydrin (DHN1 gene) putative
MAPGL58 AU300863
AU300352
cytochrome P450 (At2g23180; T20D16.19) putative
MAPBG47 AU300416
cysteine proteinase precursor
coatmer putative
MAPEM96 AU300588
clathrin binding protein putative
circadian clock coupling factor ZGT
chromatin protein (SUVH1) with SET domain
MAPGR29 AU300731
chloroplast nucleoid DNA-binding protein putative
chorismate mutase, cytosolic CM2
AU300660
MAPFF75
Putative identity / Homolog
MAPEM08 AU300550
Accession
Clone
Table 1. Continued
Metabolism
Responsive
Responsive
Responsive
Protein regulation
Protein regulation
Protein regulation
Chloroplast
Protein regulation
Protein regulation
Protein regulation
Protein regulation
Growth regulation
Metabolism
Metabolism
Cell structure
Metabolism
Chloroplast
first
secondary
stress
defense
defense
proteinase
proteinase
proteinase
folding
trafficking
trafficking
trafficking
light & circadian
secondary
secondary
chromatin
secondary
nuclear protein
second
Category of Homolog
AY042880
AJ300524
D12718
AF370589
AF417109
Z47793
AF283536
M73744
AF198627
AF348588
AY090955
AY074304
AF368237
U63534
AF255014
AF344444
AJ242648
AY080874
Accession of
Homolog
Arabidopsis thaliana
Populus euramericana
Candida maltosa
Arabidopsis thaliana
Carica candamarcensis
Citrus sinensis
Citrus x paradisi
Pisum sativum
Glycine max
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Nicotiana tabacum
Fragaria x ananassa
Citrus sinensis
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Organism
988
751
414
592
857
3006
2192
561
673
1044
1458
682
482
1493
1735
807
460
799
opt
81.1
73.1
69.5
68.3
73.2
96.0
98.0
76.0
75.5
75.3
82.9
73.1
69.7
68.9
98.1
67.7
69.4
65.1
Identity
(%)
302
323
117
293
340
651
454
204
245
384
420
353
208
730
359
393
209
455
Over
lap
132
Bull. Natl. Inst. Fruit Tree Sci. 2. 2003
AU300499
MAPEF04
dynamin-like protein SDL12
early nodulin ENOD18
elongation factor 1 alpha subunit
elongation factor fusA
endomembrane protein EMP70 precusor isolog putative
expansin 2
expansin 3
γ-adaptin 1
germin putative
MAPAE24 AU300370
MAPEM43 AU300816
MAPEM18 AU300554
MAPEM71 AU300576
AU300800
AU300591
AU300764
AU300614
MAP9C52
MAPF111
MAPH112
MAPF171
MAPGR18 AU300728
MAPBG04 AU300909
MAPEF40
glutaredoxin (Cl31)
MAPEI54
AU300826
glutamine synthetase
ent-kaurenoic acid hydroxylase (KAO2)
MAPBG88 AU300429
AU300514
dTDP-glucose 4-6-dehydratases putative
MAPBG29 AU300411
enolase
dormancy associated protein
MAPDR18 AU300468
DnaJ-1
disease responsive SOR1 putative
AU300840
MAPAT72
Putative identity / Homolog
disease resistance-like protein GS0-1
Accession
MAPGR75 AU300813
Clone
Table 1. Continued
first
Metabolism
Metabolism
Responsive
Protein regulation
Growth regulation
Growth regulation
Signal
Metabolism
Membrane
Translation
Translation
Growth regulation
Cell structure
Metabolism
Growth regulation
Protein regulation
Responsive
Responsive
second
energy
amino acid
stress
trafficking
expansin
expansin
hormone(GA)
sugar
membrane protein
elongation
elongation
nodulation
skelton
carbohydrate
dormancy
folding
defense
defense
Category of Homolog
AF406809
AF003197
AF417299
AF061286
AF527800
AF297522
AF318501
Z28386
AY063907
X71439
AJ223969
AJ271816
U25547
Z49239
AF053746
X67695
AF370296
AF403250
Accession of
Homolog
Tilia platyphyllos
Hevea brasiliensis
Castanea sativa
Arabidopsis thaliana
Malus x domestica
Prunus avium
Arabidopsis thaliana
Ricinus communis
Arabidopsis thaliana
Glycine max
Malus domestica
Vicia faba
Glycine max
Arabidopsis thaliana
Arabidopsis thaliana
Cucumis sativus
Arabidopsis thaliana
Glycine max
Organism
675
1067
700
1239
1733
1093
532
1232
1360
1450
1721
1152
776
551
803
996
960
420
opt
77.3
80.4
70.9
78.8
79.7
78.3
62.7
77.5
80.9
81.9
86.1
70.7
81.5
75.5
70.0
78.6
78.9
67.1
Identity
(%)
291
328
302
402
713
359
332
427
413
425
460
501
232
208
373
336
327
213
Over
lap
FUJII et al.: Expressed Sequence Tags from Citrus Albedo at the Initiation Stage of Rind Peeling
133
glutathione-conjugate transporter
glyoxalase I (lactoyl-glutathione lyase)
GTP-binding protein
MAPDR74 AU300898
MAP9C03
heat shock protein 70 kD
heterochromatin protein LHP1
histidine kinase receptor
MAPGR92 AU300757
MAPDD13 AU300433
AU300659
AU300850
MAPFF71
MAP9C12
histone H4
AU300618
AU300590
MAPF179
MAPF109
hormone responsive Nt-iaa28 deduced protein
AU300893
AU300810
MAPF155
MAPF147
hormone responsive-ADR11
hormone responsive CS-IAA2
MAPEM95 AU300907
HMG beta 1 protein
histone H1 putative
MAPAE25 AU300371
histone acetyltransferase
guanine nucleotide regulatory protein (ran-related GTP-binding
protein)
MAPGL01 AU300831
AU300311
glycine-rich protein LeGRP1
MAPAE88 AU300913
defense/detox
stress
protein receptor
Responsive
Responsive
Responsive
Protein
egulation
hormone(auxin)
hormone(auxin)
hormone(auxin)
folding
Cell structure chromatin
Cell structure chromatin
Cell structure chromatin
Signal
Cell structure chromatin
Responsive
Transcription G-protein
Transcription G-protein
Responsive
second
amino acid
energy
defense
defense
Cell structure cell wall
glycine decarboxylase P protein, a part of glycine cleavage complex Metabolism
MAPGL66 AU300702
Metabolism
glyceraldehyde 3-phosphate dehydrogenase protein putative
Responsive
Responsive
first
Category of Homolog
MAPDD53 AU300912
AU300600
MAPF136
Putative identity / Homolog
glutathione S-transferase/peroxidase (BI-GST/GPX)
Accession
MAPGL46 AU300691
Clone
Table 1. Continued
X69640
AF123508
AB026822
Y14072
Z79638
AY045797
AY099684
AJ298990
AF387639
X99197
Z24678
D12542
AJ224520
AY026037
X59773
AF348583
AJ002584
AF193439
Accession of
Homolog
opt
Glycine max
Nicotiana tabacum
Cucumis sativus
Arabidopsis thaliana
Sesbania rostrata
Arabidopsis thaliana
Arabidopsis thaliana
Fagus sylvatica
Arabidopsis thaliana
Medicago sativa
Vicia faba
Pisum sativum
Cicer arietinum
Lycopersicon esculentum
Pisum sativum
Arabidopsis thaliana
Arabidopsis thaliana
1028
563
618
1119
1066
695
875
1216
450
818
1643
636
1796
459
1403
579
567
Lycopersicon esculentum 1707
Organism
68.8
92.5
64.2
74.5
78.9
62.4
78.3
78.3
78.2
77.7
84.4
77.7
75.5
65.8
83.1
64.6
71.5
73.8
Identity
(%)
535
133
402
345
374
481
286
410
147
283
469
216
664
231
403
381
228
660
Over
lap
134
Bull. Natl. Inst. Fruit Tree Sci. 2. 2003
imbibition protein putative
leucine rich repeat
AU300616
AU300814
AU300832
MAPF173
MAPEI53
MAPH151
methionine synthase
AU300544
AU300776
MAPEI88
MAPH157
monodehydroascorbate reductase
MAPF133
myb-related transcription factor
MAPF134
MAPGR02 AU300720
nitrilase associated protein NAP16kDa
myb-related protein putative
MAPAE74 AU300388
AU300598
myb-like DNA-binding protein
MAPBG16 AU300847
AU300597
mitochondrial F0 ATP synthase D chain
MAPDD55 AU300448
miraculin-like protein
metallothionein-like protein
MAPGL03 AU300669
membrane protein RER1 related to ER retrieval
lipoxygenase(loxC)
Lea5 protein
MAPEM68 AU300574
AU300885
laminin receptor-like protein (40S ribosomal protein SA)
AU300350
MAP9U75
IAA-amino acid hydrolase
MAP9C10
AU300779
MAPH165
hormone responsive GAI-like protein 1 (GAI1)
importin alpha (LeKAP alpha)
AU300921
MAPAT85
Putative identity / Homolog
MAPEM79 AU300821
Accession
Clone
Table 1. Continued
vitamin
ATPase
proteinase
amino acid
stress
trafficking
defence
protein receptor
stress
ribosomal
transport (nuclear)
disease response
stress
hormone
hormone (IAA)
Transcription myb
Transcription myb
Signal
second
hormone
Transcription myb
Metabolism
Membrane
Protein
regulation
Metabolism
Responsive
Protein
regulation
Responsive
Signal
Responsive
Translation
Membrane
Responsive
Signal
Responsive
first
Category of Homolog
Z96936
AF161711
AY063912
AJ251686
U06461
AJ271469
AF283533
X83499
AF320905
AB010946
U37839
Y07748
Z46824
U01955
AF017252
AJ271668
AY074380
AF378125
Accession of
Homolog
620
537
407
opt
1505
1845
1113
1006
522
1312
1370
3678
723
1916
776
Arabidopsis thaliana
476
Pimpinella brachycarpa 1332
Arabidopsis thaliana
Catharanthus roseus
Pisum sativum
Arabidopsis thaliana
Citrus x paradisi
Catharanthus roseus
Citrus unshiu
Arabidopsis thaliana
Lycopersicon esculentum 529
Oryza sativa
Citrus sinensis
Arabidopsis thaliana
Lycopersicon esculentum 719
Cicer arietinum
Arabidopsis thaliana
Vitis vinifera
Organism
66.4
67.6
74.1
70.0
78.3
77.3
93.4
77.1
98.0
73.2
61.0
75.6
95.4
77.0
77.3
67.1
69.5
67.8
Identity
(%)
265
735
401
243
461
481
843
258
400
295
419
553
417
381
238
301
249
199
Over
lap
FUJII et al.: Expressed Sequence Tags from Citrus Albedo at the Initiation Stage of Rind Peeling
135
nucleoside diphosphate kinase
nucleotide sugar epimerase-like protein (F14O13.1)
MAPEM15 AU300553
AU300619
AU300310
AU300905
AU300664
AU300336
MAPF183
MAP9C02
MAPAT89
MAPFF81
MAP9U30
pectin acetylesterase
pectinesterase (PECS-1.1)
permease, possibly related to nucleobase-ascorbate transporter
peroxidase
MAPEM60 AU300570
MAPGR46 AU300738
MAPEM84 AU300584
MAPFF65
peroxidase isozyme 40K precursor cationic
phosphate/triose-phosphate translocator TPT precursor, non-green
plastid
phosphoenolpyruvate carboxylase (pepc)
MAPEM25 AU300825
MAPGL85 AU300712
MAPEI74
plasma membrane intrinsic protein (SIMIP)
MAPFF85
AU300665
photosystem II type I chlorophyll a/b binding protein
MAPDR88 AU300492
AU300541
peroxidase ascorbate cytosolic
MAPGL16 AU300676
AU300656
pectin methylesterase
MAPAE10 AU300361
pectate lyase
pathogenesis-related protein PR1 (Ypr1)
pathogenesis-related protein 5
O-acetylserine (thiol) lyase B
nuclease (nucZe2) bifunctional
AU300829
MAPFF09
Putative identity / Homolog
Accession
Clone
Table 1. Continued
defense
defense
amino acid (cystein)
nucleic acid
nucleic acid
Membrane
Metabolism
Metabolism
Metabolism
Responsive
Responsive
Responsive
membrane protein
photosystem
energy
sugar
stress
stress
stress
transporter
Cell structure cell wall
Cell structure cell wall
Cell structure cell wall
Membrane
second
senescence
Cell structure cell wall
Responsive
Responsive
Metabolism
Metabolism
Metabolism
Growth
regulation
first
Category of Homolog
AF003728
X64459
AF512995
U13632
D42065
AF022213
L08199
AY093137
U82973
X99348
Z71753
AB048260
U64806
AF154636
AJ271728
AY062625
U50150
U90266
Accession of
Homolog
Arabidopsis thaliana
Arabidopsis thaliana
Citrus junos
Brassica oleracea
Nicotiana tabacum
Fragaria x ananassa
Gossypium hirsutum
Arabidopsis thaliana
Citrus sinensis
Vigna radiata
Nicotiana plumbaginifolia
Salix gilgiana
Brassica napus
Nicotiana tabacum
Arabidopsis thaliana
Arabidopsis thaliana
Glycine max
Zinnia elegans
Organism
2787
432
1520
409
475
679
1577
566
1992
717
634
653
401
1462
579
496
1451
718
opt
78.2
69.8
98.1
63.2
60.0
73.7
84.9
74.4
96.5
70.5
75.8
79.7
60.5
79.1
72.6
71.0
80.7
72.6
Identity
(%)
930
222
313
359
380
266
430
211
423
312
227
217
352
470
249
224
460
299
Over
lap
136
Bull. Natl. Inst. Fruit Tree Sci. 2. 2003
polygalacturonase isoenzyme 1 beta subunit
proline rich protein, TPRP-F1
proteasome (26S ) beta subunit
proteasome (26S) subunit 4-like protein SPH
protein kinase (fl-E6)
protein kinase Ndr kinase
MAPAE26 AU300857
MAPDR59 AU300484
MAPDD40 AU300791
MAPGL88 AU300714
MAPFF11
protein kinase Pti1 kinase-like protein (Pti1a)
protein kinase-like protein
protein phosphatase 2C putative
proteinase nClpP4 (nuclear encoded ClpP4)
psaG Photosystem I subunit V precursor
psbO mRNA for oxygen evolving enhancer protein 1 precursor
pyruvate decarboxylase
MAPGR17 AU300727
MAPGL48 AU300692
MAPAE85 AU300392
MAPDD90 AU300827
MAPDD35 AU300441
MAPGR71 AU300843
MAPGR11 AU300726
AU300630
polyubiquitin rub1
MAPGR01 AU300719
AU300612
polyubiquitin (ubq8)
AU300356
MAP9U96
poly(A)-binding protein (PABP)
MAPF167
AU300802
MAPFF19
plastid-specific ribosomal protein 3 precursor (Psrp-3)
polyubiquitin (ubq4)
AU300851
MAPFF47
Putative identity / Homolog
MAPAE05 AU300894
Accession
Clone
Table 1. Continued
mRNA stability
proteolysis
proteolysis
proteolysis
Metabolism
Metabolism
Metabolism
Protein
regulation
Protein
regulation
Protein
regulation
Responsive
Protein
regulation
Protein
regulation
Protein
regulation
Protein
regulation
energy
photosystem
photosystem
proteinase
phosphatase
PK
defense
PK
PK
proteolysis
proteolysis
Cell structure cell wall
Protein
regulation
Protein
regulation
Protein
regulation
second
ribosomal
Cell structure cell wall
Translation
Translation
first
Category of Homolog
AF193791
AB043960
AJ245630
AB022329
AY050873
AY099731
AF249317
AB047278
AF218378
AJ291500
D78172
X61395
X76064
L05917
U33014
U63373
AF190655
AF239218
Accession of
Homolog
Fragaria x ananassa
Bruguiera gymnorrhiza
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Glycine max
Arabidopsis thaliana
Gossypium hirsutum
Brassica napus
Spinacia oleracea
Lycopersicon esculentum
Oryza sativa
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Nicotiana tabacum
Spinacia oleracea
Organism
1866
797
1202
674
906
433
923
496
416
949
591
739
1633
840
1511
585
1327
898
opt
81.9
78.5
70.3
64.7
73.0
65.0
80.8
67.4
59.3
75.6
70.8
64.1
85.7
64.8
83.4
59.8
79.1
66.4
Identity
(%)
570
265
583
394
348
254
286
239
376
356
243
473
434
486
430
423
431
533
Over
lap
FUJII et al.: Expressed Sequence Tags from Citrus Albedo at the Initiation Stage of Rind Peeling
137
AU300355
AU300908
MAP9U94
MAPFF33
receptor-like LRR protein kinase
receptor-like protein kinase (RLK)
receptor-like protein kinase (WRK)
ribonucleoprotein-like
ribosomal protein small subunit 4e
MAPEM04 AU300849
AU300397
MAPGR89 AU300790
MAPGR49 AU300740
AU300516
AU300638
AU300611
AU300639
MAPAT67
MAPEF47
MAPFF27
MAPF166
MAPFF29
ribosomal (50S) protein L29 putative
ribosomal (60S) protein (rib 60S)
ribosomal (60S) protein L2
MAPAE13 AU300363
MAPDR10 AU300465
AU300663
AU300649
MAPFF80
MAPFF50
ribosomal (60S) protein L37 putative
ribosomal (50S) protein L28 chloroplast putative
MAPAE78 AU300390
ribosomal (40S) protein S5 putative
ribosomal (40S) protein S15 (Sb23) probable
ribosomal (40S) protein
receptor-kinase isolog putative
MAPDR38 AU300478
RD2 protein, responsive to desiccation
ran GTPase activating protein
raffinose synthase
AU300540
MAPEI72
Putative identity / Homolog
QM family gene (60S ribosomal protein L10)
Accession
MAPEM48 AU300803
Clone
Table 1. Continued
second
sugar
ribosomal
Translation
Translation
Translation
Translation
Translation
Translation
Translation
Translation
Translation
Group protein
Responsive
Signal
Signal
Signal
Responsive
ribosomal
ribosomal
ribosomal
ribosomal
ribosomal
ribosomal
ribosomal
ribosomal
ribosomal
defense
protein receptor
protein receptor
protein receptor
stress
Transcription nuclear import
Metabolism
Translation
first
Category of Homolog
AF370216
X62500
AJ278460
AY045966
AY072373
AY045846
AF051217
AY054622
X79300
AY136466
AB073628
AF149037
M84660
AY056128
AB039925
AF215731
E25448
AB001891
Accession of
Homolog
Arabidopsis thaliana
Nicotiana tabacum
Juglans regia
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Picea mariana
Arabidopsis thaliana
Gossypium hirsutum
Arabidopsis thaliana
Nicotiana tabacum
Prunus dulcis
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Medicago sativa
Glycine max
Solanum melongena
Organism
847
1154
994
960
595
960
1319
718
1404
749
1762
861
630
854
544
770
984
2124
opt
81.4
77.7
71.2
62.8
68.6
72.1
80.7
79.6
84.0
63.9
71.5
79.2
62.3
73.9
72.4
68.7
76.4
80.6
Identity
(%)
296
382
425
651
296
408
436
230
393
451
758
288
477
322
221
364
348
654
Over
lap
138
Bull. Natl. Inst. Fruit Tree Sci. 2. 2003
ribosomal protein L17-1
ribosomal protein L26 like
ribosomal protein L5 putative
ribosomal protein S14 (rps14)
ribosomal protein S24 40S
ribosomal protein S6 (rps6 gene)
ribosomal protein S8
ribosomal protein L23
RING zinc finger ankyrin protein putative
MAPBG72 AU300424
MAPGR47 AU300739
MAPAE16 AU300365
MAPEM51 AU300563
AU300340
AU300332
AU300601
AU300631
AU300793
AU300819
AU300782
MAP9U10
MAPEM64 AU300571
AU300805
MAP9U44
MAPF107
MAPF139
MAPFF15
MAP9C42
MAP9C55
MAPH176
serine carboxypeptidase precursor (SCP) putative
MAPAE80 AU300391
Protein
regulation
Metabolism
Metabolism
Translation
proteinase
energy
energy
energy
ribosomal
Transcription RNA helicase
S-adenosyl-L-methionine decarboxylase proenzyme
S-adenosyl-methinonine synthetase
ribosomal
ribosomal
ribosomal
ribosomal
ribosomal
ribosomal
ribosomal
ribosomal
ribosomal
Transcription RNA helicase
Metabolism
MAPGL26 AU300683
second
ribosomal
Transcription zinc finger
Translation
Translation
Translation
Translation
Translation
Translation
Translation
Translation
Translation
Translation
first
Category of Homolog
S-adenosyl-homocysteine hydrolase (SHH)
RSP ribosome sedimenting protein
RNA helicase ATP-dependent putative
RNA helicase DRH1
ribosomal protein L19
MAPGL62 AU300700
AU300326
MAP9C49
Putative identity / Homolog
ribosomal protein L17 (rpl17)
Accession
MAPGR28 AU300730
Clone
Table 1. Continued
AY072822
AJ277206
U12573
M81885
AB021873
AY091050
AY062591
AY042842
AF349961
U64436
AJ277533
AF298769
AF026079
AY056194
D78495
Z31720
AF264022
AF034948
Accession of
Homolog
Gossypium hirsutum
Camellia sinensis
Catharanthus roseus
Petroselinum crispum
Pisum sativum
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Daucus carota
Zea mays
Asparagus officinalis
Zea mays
Lupinus luteus
Arabidopsis thaliana
Brassica rapa
Nicotiana tabacum
Poa secunda
Zea mays
Organism
948
2486
629
555
482
614
983
540
900
827
2395
1147
1423
441
1295
2095
1447
805
opt
71.6
82.6
84.3
78.3
61.9
68.5
76.3
67.4
74.9
70.6
76.6
69.7
78.3
72.5
72.5
76.5
69.5
77.3
Identity
(%)
405
748
178
198
391
289
334
258
351
361
824
554
425
171
523
761
656
269
Over
lap
FUJII et al.: Expressed Sequence Tags from Citrus Albedo at the Initiation Stage of Rind Peeling
139
AU300806
AU300667
AU300920
MAP9U71
MAPFF87
MAPF178
small GTP-binding protein
small GTP-binding protein rab-type
splicing factor (ORF1) putative
MAPAE58 AU300384
MAPGL10 AU300673
AU300313
AU300325
AU300836
AU300402
AU300903
AU300645
MAP9C07
MAP9C48
MAPFF18
MAPAT92
MAPF104
MAPFF42
AU300789
MAPH194
MAPGR74 AU300750
suppressor of K+ transport growth defect-like protein (SKD1)
AU300617
MAPF176
TFIIH p44/SSL1-like protein
synaptobrevin-related protein (SAR1)
sucrose synthase CitSUSA
MAPGL64 AU300701
subtilisin serine protease putative
stress responsive-SRG1
stress responsive phi-1 protein putative
stearoyl-acyl-carrier protein desaturase
starch synthase granule-bound
sinapyl alcohol dehydrogenase
AU300844 σfactor SigB
SF16 pollen specific gene putative
serine/threonine protein kinase-like protein
serine protease
serine palmitoyltransferase
Putative identity / Homolog
MAPGL92 AU300717
MAPFF73
MAPDD42 AU300901
Accession
Clone
Table 1. Continued
Transcription
Protein
regulation
Membrane
Metabolism
Protein
regulation
Responsive
Responsive
Metabolism
Metabolism
Transcription
Transcription
Transcription
Cell structure
Transcription
Morphogenesis
Protein
regulation
Protein
regulation
Metabolism
first
TF
trafficking
transport
sugar
proteinase
stress
stress
lipid
carbohydrate
splicing
G-protein
G-protein
lignin
TF
pollen
proteinase
proteinase
amino acid
second
Category of Homolog
AF499443
M90418
AF165422
AB022091
AY051009
X79052
AF326891
M59857
AF097922
AJ299058
AB024994
Z49902
AF273256
AB019943
AY099822
AY062765
X98929
AJ242659
Accession of
Homolog
878
opt
590
895
909
1189
532
680
641
Arabidopsis thaliana
Arabidopsis thaliana
Mesembryanthemum
crystallinum
Citrus unshiu
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Ricinus communis
862
983
1408
1373
533
437
435
1287
Astragalus membranaceus 1129
Cicer arietinum
Cicer arietinum
Pisum sativum
Populus tremuloides
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Lycopersicon esculentum 1913
Solanum tuberosum
Organism
76.4
70.7
85.3
94.7
69.3
66.0
76.7
80.2
75.1
80.6
76.1
73.0
79.8
67.3
78.0
73.1
69.1
71.3
Identity
(%)
296
427
380
304
238
244
150
405
410
181
326
381
371
272
223
253
895
391
Over
lap
140
Bull. Natl. Inst. Fruit Tree Sci. 2. 2003
Translation
Growth
regulation
MAPGL73 AU300707 translational elongation factor Tu (tufA) chloroplast
AU300521 translationally controlled tumor protein (TCTP)
AU300906 transporter MATE efflux family protein putative
AU300928 triosphosphate isomerase
AU300655 ubiquitin
AU300593 ubiquitin activating enzyme E1 (ECR1) putative
MAPEF57
MAPFF10
MAPEF14
MAPFF62
MAPF115
Protein
regulation
MAPFF44
Protein
regulation
AU300604 ubiquitin/ribosomal protein 27a (Ubi)
AU300530 UDP-glucose dehydrogenase
MAPF149
MAPEI33
Metabolism
Membrane
MAPGL77 AU300824 UMP synthase (pyr5-6)
MAPDD60 AU300452 vacuolar H+-ATPase subunit E
Metabolism
Protein
regulation
MAPAE81 AU300879 ubiquitin SMT3 protein
AU300647 ubiquitin extension protein UBQ1putative
Protein
regulation
MAPGR03 AU300721 ubiquitin conjugating enzyme (UBC4)
Protein
regulation
Protein
regulation
Metabolism
Membrane
Translation
Metabolism
MAPGR60 AU300744 translation initiation factor
AU300780 transaldolase (TAL)
MAPH168
Metabolism
Metabolism
first
second
ATPase
energy
sugar
proteolysis
proteolysis
proteolysis
proteolysis
proteolysis
proteolysis
sugar
transporter
cell cycle
elongation
initiation
TF / CHS
sugar
amino acid
energy
Category of Homolog
Transcription
AU300532 threonine synthase
MAPEI40
Putative identity / Homolog
MAPGR34 AU300733 transcription factor KAP-2
AU300651 thioredoxin
Accession
MAPFF53
Clone
Table 1. Continued
AF165939
U22260
U53418
AF298826
X99609
AY035148
L29077
AF051135
X67957
J04121
AF360254
AF091455
AF234537
AF098672
AF293343
AF184164
AF082894
Z70677
Accession of
Homolog
740
1267
opt
1184
1037
Citrus limon
Nicotiana tabacum
Glycine max
Prunus avium
Arabidopsis thaliana
Arabidopsis thaliana
Pisum sativum
Arabidopsis thaliana
Arabidopsis thaliana
Coptis japonica
Arabidopsis thaliana
Hevea brasiliensis
1717
541
1200
1787
818
1298
1770
1222
1150
1161
592
1907
Pelargonium graveolens 1008
Brassica oleracea
Medicago truncatula
Lycopersicon esculentum 465
Solanum tuberosum
Ricinus communis
Organism
97.7
72.9
83.8
89.9
82.6
85.1
78.1
78.6
80.3
82.7
64.9
83.1
77.0
81.6
77.6
73.9
69.9
80.9
Identity
(%)
356
218
339
436
235
397
603
392
356
336
353
568
378
358
340
180
346
404
Over
lap
FUJII et al.: Expressed Sequence Tags from Citrus Albedo at the Initiation Stage of Rind Peeling
141
Cell structure cell wall
MAP9C39
Transcription
Transcription
MAPAE93 AU300919 zinc-finger C2H2 protein SERRATE (SE)
MAPAE17 AU300817 zinc-finger protein Lsd1 negative regulator to cell death
AU300320 xyloglucan endotransglycosylase-related protein XTR-7 putative
Membrane
zinc finger
zinc finger
membrane protein
elongation
MAPEM39 AU300558 water channel protein
second
transporter
Translation
first
Category of Homolog
MAPGL74 AU300708 vitronectin-like adhesion protein (elongation factor-1 alpha)
Putative identity / Homolog
Membrane
Accession
MAPGL06 AU300671 vacuolar H+-pyrophosphatase (vp1)
Clone
Table 1. Continued
U87833
AF311221
U43489
U73467
U04632
AF367446
Accession of
Homolog
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Mesembryanthemum
crystallinum
Nicotiana tabacum
Prunus persica
Organism
552
605
530
1220
734
1495
opt
63.8
69.0
65.8
77.8
71.9
85.4
Identity
(%)
345
277
310
427
334
431
Over
lap
142
Bull. Natl. Inst. Fruit Tree Sci. 2. 2003
FUJII et al.: Expressed Sequence Tags from Citrus Albedo at the Initiation Stage of Rind Peeling
143
Table 2. Functional classification of C. unshiu fruit ESTs from albedo at initiation stage of rind peeling. The ESTs that had
sequence similarity to known genes were classified based on their biological functions.
Classification based on biological function
Metabolism
Photo/electron
Sugar/carbohydrate
Amino acid
Nucleic acid
Vitamin
Lipid
Secondary
Cell structure
Cell wall
Skelton
Lignin
Chromatin
Membrane
Transport
ATPases
Membrane protein
Growth regulation
Cell cycle
Nodulation
Expansin
Dormancy
Light & circadian
Senescence
Morphogenesis
Pollen
Signaling
Calcium
Hormone synthesis
Protein receptor
Number of
Number of
ESTs with
ESTs with
Classification based on biological function
matches
matches
(non(redundant)
redundant)
55
16
14
6
4
3
4
8
36
20
5
3
8
21
7
2
12
25
6
1
7
9
1
1
1
1
12
4
3
5
59
16
16
6
4
3
4
10
39
21
5
4
9
26
7
2
17
38
10
1
9
16
1
1
1
1
12
4
3
5
Responsive
Defense
Hormone
Stress
Transcription
Zinc finger
G-protein
Myb
TF
Helicase
Nuclear transport
Splicing
Translation
Initiation factor
Elongation
Ribosomal proteins
mRNA stability
Protein regulation
Folding
PK
Phosphatase
Proteinase
Proteolysis
Trafficking
Chloroplast protein
unclassified
Total
Number of
ESTs with
matches
(nonredundant)
Number of
ESTs with
matches
(redundant)
68
14
5
49
19
4
5
3
3
2
1
1
36
2
6
27
1
53
7
3
1
17
15
10
95
15
7
73
19
4
5
3
3
2
1
1
43
3
8
30
2
61
8
3
1
21
16
12
2
8
2
8
336
403
Bull.
果樹研究所研究報告 第
Natl. Inst. Fruit Tree 2Sci.
号 2003
2. 2003
144
ウンシュウミカン果実の剥皮開始期における
アルベドに由来する EST の解析
藤井 浩・喜多正幸†1・島田武彦・遠藤朋子・大村三男
独立行政法人農業技術研究機構
果樹研究所カンキツ研究部興津
424-0292 静岡県清水市
では他の生物の EST と相同性があり，
25 クローン
（4.0％）
摘 要
についてはシロイヌナズナのゲノム塩基配列とのみ相同
性があった．残りの 175 クローン（28.0％）については
ウンシュウミカン果実の剥皮開始期アルベドで発現し
他の生物の塩基配列との相同性は見られなかった．機能
た遺伝子から作成した cDNA ライブラリーについて，ラ
が推定されたクローンのうち 222 クローンについて，機
ンダムに cDNA を選択してシーケンスし，626 の cDNA
能推定の結果を一覧表に示した．この cDNA ライブラ
クローンの EST を得た．相同性検索による EST の機能
リーの特徴として，細胞壁にかかわる遺伝子が多くみら
解析の結果，336 クローン（53.7％）で遺伝子機能が推定
れた．
された．残りのクローンのうち，90 クローン（14.3％）
†1
現 果樹研究所生理機能部
305-8605 茨城県つくば市