五、計畫成果自評 目標：
C. sorokiniana plastid DNA small subunit (16S-like) ribosomal RNA Length=1493
Score = 835 bits (421), Expect = 0.0 Identities = 443/451 (98%), Gaps = 0/451 (0%) Strand=Plus/Plus
2
Chlorella pyrenoidosa 16S rRNA gene CS12 region, strain IAM C-210 Length=450
Score = 817 bits (412), Expect = 0.0 Identities = 436/444 (98%), Gaps = 0/444 (0%) Strand=Plus/Plus
3
Chlorella sp. 16S rRNA gene CS12 region, isolate V Length=450
Score = 811 bits (409), Expect = 0.0 Identities = 433/441 (98%), Gaps = 0/441 (0%) Strand=Plus/Plus
4
Chlorella vulgaris (strain 211-1e) chloroplast gene for 16S ribosomal RNA Length=1450
Score = 809 bits (408), Expect = 0.0 Identities = 442/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
5
Chlorella vulgaris chloroplast DNA for small subunit ribosomal RNA Length=1494
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
6
Auxenochlorella protothecoides 16S small subunit ribosomal RNA gene, partial sequence; chloroplast
Length=1475
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Minus
7
Chlorella vulgaris C-27 chloroplast DNA, complete sequence Length=150613
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
8
Chlorella vulgaris 16S ribosomal RNA gene, partial sequence; chloroplast gene for chloroplast product
Length=578
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
9
Chlorella sp. 16S rRNA gene CS12 region, isolate 21 Length=464
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
10
Chlorella sp. 16S rRNA gene CS12 region, isolate 216 Length=464
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
11
Chlorella vulgaris chloroplast 16S rRNA gene, partial, strain NIES 227 Length=479
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
12
Chlorella vulgaris chloroplast 16S rRNA gene, partial, strain IAM C-27 Length=475
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
13
Chlorella vulgaris chloroplast 16S rRNA gene, partial, strain CCAP 211/19 Length=481
Score = 793 bits (400), Expect = 0.0 Identities = 440/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
14
Chlorella kessleri (strain 211-11h) chloroplast gene for 16S ribosomal RNA Length=1545
Score = 767 bits (387), Expect = 0.0 Identities = 436/451 (96%), Gaps = 1/451 (0%) Strand=Plus/Plus
15
Chlorella kessleri plastid DNA for 16S-like rRNA small subunit, strain SAG 211-11g
Length=1592
Score = 767 bits (387), Expect = 0.0 Identities = 436/451 (96%), Gaps = 1/451 (0%) Strand=Plus/Plus
表3-4 BG33 以 CS34 放大序列在 NCBI 進行比對資訊 項
目 以NCBI 中 Blast 所得之資訊
1
C.sorokiniana plastid DNA small subunit (16S-like) ribosomal RNA Length=1493
Score = 835 bits (421), Expect = 0.0 Identities = 443/451 (98%), Gaps = 0/451 (0%) Strand=Plus/Plus
2
Chlorella pyrenoidosa 16S rRNA gene CS12 region, strain IAM C-210 Length=450
Score = 817 bits (412), Expect = 0.0 Identities = 436/444 (98%), Gaps = 0/444 (0%) Strand=Plus/Plus
3
Chlorella sp. 16S rRNA gene CS12 region, isolate V Length=450
Score = 811 bits (409), Expect = 0.0 Identities = 433/441 (98%), Gaps = 0/441 (0%) Strand=Plus/Plus
4
Chlorella vulgaris (strain 211-1e) chloroplast gene for 16S ribosomal RNA Length=1450
Score = 809 bits (408), Expect = 0.0 Identities = 442/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
5
Chlorella vulgaris chloroplast DNA for small subunit ribosomal RNA Length=1494
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
6
Auxenochlorella protothecoides 16S small subunit ribosomal RNA gene, partial sequence; chloroplast
Length=1475
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Minus
7
Chlorella vulgaris C-27 chloroplast DNA, complete sequence Length=150613
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
8
Chlorella vulgaris 16S ribosomal RNA gene, partial sequence; chloroplast gene for chloroplast product
Length=578
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
9
Chlorella sp. 16S rRNA gene CS12 region, isolate 21 Length=464
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%)
10
Chlorella sp. 16S rRNA gene CS12 region, isolate 216 Length=464
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
11
Chlorella vulgaris chloroplast 16S rRNA gene, partial, strain NIES 227 Length=479
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
12
Chlorella vulgaris chloroplast 16S rRNA gene, partial, strain IAM C-27 Length=475
Score = 801 bits (404), Expect = 0.0 Identities = 441/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
13
Chlorella vulgaris chloroplast 16S rRNA gene, partial, strain CCAP 211/19 Length=481
Score = 793 bits (400), Expect = 0.0 Identities = 440/452 (97%), Gaps = 1/452 (0%) Strand=Plus/Plus
14
Chlorella kessleri (strain 211-11h) chloroplast gene for 16S ribosomal RNA Length=1545
Score = 767 bits (387), Expect = 0.0 Identities = 436/451 (96%), Gaps = 1/451 (0%) Strand=Plus/Plus
15
Chlorella kessleri plastid DNA for 16S-like rRNA small subunit, strain SAG 211-11g
Length=1592
Score = 767 bits (387), Expect = 0.0 Identities = 436/451 (96%), Gaps = 1/451 (0%) Strand=Plus/Plus
表3-5 BG33 以 NS12 放大序列 NCBI 進行比對資訊 項
目 以NCBI 中 Blast 所得之資訊
1
Chlorella sp. Mary 9/21 BT-10w 18S ribosomal RNA gene, partial sequence Length=1795
Score = 1051 bits (530), Expect = 0.0
Identities = 530/530 (100%), Gaps = 0/530 (0%) Strand=Plus/Plus
2
Didymogenes palatina 18S ribosomal RNA gene, partial sequence Length=1769
Score = 1051 bits (530), Expect = 0.0
Identities = 530/530 (100%), Gaps = 0/530 (0%) Strand=Plus/Plus
3
Diacanthos belenophorus 18S ribosomal RNA gene, partial sequence Length=1771
Score = 1043 bits (526), Expect = 0.0
Identities = 529/530 (99%), Gaps = 0/530 (0%) Strand=Plus/Plus
4
Chlorella sp. NDem 9/21 T-13d 18S ribosomal RNA gene, partial sequence Length=1759
Score = 1037 bits (523), Expect = 0.0
Identities = 530/531 (99%), Gaps = 1/531 (0%) Strand=Plus/Plus
5
Chlorella sp. MDL4-1 18S ribosomal RNA gene, partial sequence Length=1796
Score = 1037 bits (523), Expect = 0.0
Identities = 530/531 (99%), Gaps = 1/531 (0%) Strand=Plus/Plus
6
Chlorella sp. JL 2/4 18S ribosomal RNA gene, partial sequence Length=1796
Score = 1037 bits (523), Expect = 0.0
Identities = 530/531 (99%), Gaps = 1/531 (0%) Strand=Plus/Plus
7
Chlorella pyrenoidosa gene for 18S rRNA, partial sequence Length=2162
Score = 1037 bits (523), Expect = 0.0
Identities = 530/531 (99%), Gaps = 1/531 (0%) Strand=Plus/Plus
8
Chlorella sp. MBIC10595 gene for 18S rRNA, partial sequence Length=1752
Score = 1037 bits (523), Expect = 0.0
Identities = 530/531 (99%), Gaps = 1/531 (0%) Strand=Plus/Plus
9
Chlorella vulgaris strain CCAP 211/11F 18S ribosomal RNA gene, complete sequence
Length=1777
Score = 1037 bits (523), Expect = 0.0
Identities = 530/531 (99%), Gaps = 1/531 (0%) Strand=Plus/Plus
10
Chlorella sp. (Ssh) gene for ribosomal RNA, small subunit Length=1797
Score = 1037 bits (523), Expect = 0.0
Identities = 530/531 (99%), Gaps = 1/531 (0%) Strand=Plus/Plus
11
Chlorella sp. (Esh) gene for ribosomal RNA, small subunit Length=1797
Score = 1037 bits (523), Expect = 0.0
Identities = 530/531 (99%), Gaps = 1/531 (0%) Strand=Plus/Plus
12
Chlorella sp. AN 1-3 18S ribosomal RNA gene, partial sequence Length=2125
Score = 1035 bits (522), Expect = 0.0
Identities = 528/530 (99%), Gaps = 0/530 (0%) Strand=Plus/Plus
13
Chlorella lobophora 18S rRNA gene, strain Andreyeva 750-I Length=1796
Score = 1031 bits (520), Expect = 0.0
Identities = 527/530 (99%), Gaps = 0/530 (0%) Strand=Plus/Plus
14
Chlorella sp. MRBG1 genes for 18S rRNA, internal transcribed spacer 1, 5.8S rRNA, internal transcribed spacer 2, partial and complete sequence
Length=3948
Score = 1029 bits (519), Expect = 0.0
Identities = 529/531 (99%), Gaps = 1/531 (0%) Strand=Plus/Plus
15
Chlorella vulgaris genes for 18S rRNA, ITS1, 5.8S rRNA, strain:takaP-3-A2 Length=3565
Score = 1029 bits (519), Expect = 0.0
Identities = 529/531 (99%), Gaps = 1/531 (0%) Strand=Plus/Plus
表3-6 MN41 以 18S12 放大序列在 NCBI 進行比對資訊 項
目 以NCBI 中 Blast 所得之資訊
1
Picochlorum sp. UTEX 2378 18S small subunit ribosomal RNA gene, partial sequence
Length=1692
Score = 1758 bits (887), Expect = 0.0
Identities = 934/940 (99%), Gaps = 2/940 (0%) Strand=Plus/Plus
2
Nanochlorum sp. MBIC10208 gene for 18S rRNA, partial sequence Length=1735
Score = 1758 bits (887), Expect = 0.0
Identities = 934/940 (99%), Gaps = 2/940 (0%) Strand=Plus/Plus
3
Picochlorum sp. UTEX 2491 18S small subunit ribosomal RNA gene, partial sequence
Length=1692
Score = 1742 bits (879), Expect = 0.0
Identities = 932/940 (99%), Gaps = 2/940 (0%) Strand=Plus/Plus
4
Picochlorum oklahomensis 18S small subunit ribosomal RNA gene, partial sequence
Length=1692
Score = 1742 bits (879), Expect = 0.0
Identities = 932/940 (99%), Gaps = 2/940 (0%) Strand=Plus/Plus
5
Nannochloris sp. RCC 011 18S rRNA gene Length=1792
Score = 1742 bits (879), Expect = 0.0
Identities = 932/940 (99%), Gaps = 2/940 (0%) Strand=Plus/Plus
6
Nannochloris maculata gene for 18S rRNA, partial sequence Length=1788
Score = 1727 bits (871), Expect = 0.0
Identities = 930/940 (98%), Gaps = 2/940 (0%) Strand=Plus/Plus
7
Picochlorum sp. RCC115 18S ribosomal RNA gene, partial sequence Length=1737
Score = 1727 bits (871), Expect = 0.0
Identities = 931/940 (99%), Gaps = 3/940 (0%) Strand=Plus/Plus
8
Nannochloris atomus gene for 18S rRNA, partial sequence, strain:CCAP 251/7 Length=1788
Score = 1719 bits (867), Expect = 0.0
Identities = 929/940 (98%), Gaps = 2/940 (0%) Strand=Plus/Plus
9
Nannochloris sp. MBIC10596 gene for 18S rRNA, partial sequence, strain:
MBIC10596 Length=1746
Strand=Plus/Plus
10
Nanochlorum sp. MBIC10053 gene for 18S rRNA, partial sequence Length=1749
Score = 1679 bits (847), Expect = 0.0
Identities = 924/940 (98%), Gaps = 2/940 (0%) Strand=Plus/Plus
11
Prasinoderma sp. MBIC10059 gene for 18S rRNA, partial sequence, strain:
MBIC10059 Length=1743
Score = 1679 bits (847), Expect = 0.0
Identities = 924/940 (98%), Gaps = 2/940 (0%) Strand=Plus/Plus
12
Nanochlorum sp. MBIC10091 gene for 18S rRNA, partial sequence Length=1749
Score = 1663 bits (839), Expect = 0.0
Identities = 922/940 (98%), Gaps = 2/940 (0%) Strand=Plus/Plus
13
Nannochloris sp. AS 2-10 18S ribosomal RNA gene, partial sequence Length=1794
Score = 1655 bits (835), Expect = 0.0
Identities = 919/940 (97%), Gaps = 1/940 (0%) Strand=Plus/Plus
14
Chlorella sp. 18S rRNA gene, isolate Yanaqocha RA1 Length=1794
Score = 1655 bits (835), Expect = 0.0
Identities = 918/940 (97%), Gaps = 1/940 (0%) Strand=Plus/Plus
15
Picochlorum oculatum 18S small subunit ribosomal RNA gene, partial sequence
Length=1692
Score = 1647 bits (831), Expect = 0.0
Identities = 920/940 (97%), Gaps = 2/940 (0%) Strand=Plus/Plus
圖1-1 物理化學吸收及生物固定二氧化碳整合系統構想圖
圖1-2 CA酵素在微藻光合作用中所扮演之角色
0 50 100
4 5 6 7 8 9 10 11 12 13
% of total CO 2
pH
CO3
HCO 2-3
CO -2
圖1-3 不同pH下之碳酸物種分佈圖
圖3-1 選擇性培養基篩選成果
圖3-2 分離藻株之SEM圖像 5%植種量，
生長2 週 5%繼代，生長 2 週
36 管/39 管
36 管每管劃碟 2-3 盤 共100 盤，生長 2 週 73 盤明顯生長，選定
29 株
藻源：海水 (興達港) 培養基種類：四種 酸鹼值：pH 2~5 碳酸鎂濃度：0.1~1g/L
29 株植種 80 管
5%繼代 50 瓶 每株1-2 瓶
圖3-3 BG33以16S12放大DNA產物之梯度電泳圖
圖3-4 BG33以CS12、CS34、NS12放大DNA產物之電泳圖 1500 bP
2000 bp
marker marker
marker marker
600 bp 500 bp
圖3-5 MN41以18S12放大DNA產物之梯度電泳圖 1000 bp
1500 bp
marker marker
圖3-6 BG33以16S12放大之定序光譜(正向)
圖3-6 BG33以16S12放大之定序光譜(正向)(續)
圖3-7 BG33以16S12放大之定序光譜(反向)
圖3-7 BG33以16S12放大之定序光譜(反向)(續)
圖3-8 BG33以CS12放大之定序光譜(正向)
圖3-9 BG33以CS12放大之定序光譜(反向)
圖3-10 BG33以CS34放大之定序光譜(正向)
圖3-11 BG33以CS34放大之定序光譜(反向)
圖3-12 BG33以NS12放大之定序光譜(正向)
圖3-13 BG33以NS12放大之定序光譜(反向)
圖3-14 MN41以18S12放大之定序光譜(正向)
圖3 -14 MN41以18S12放大之定序光譜(正向)(續)
圖3-15 MN41以18S12放大之定序光譜(反向)
gi|4868347/1-1441
BG CS12/1-709
gi|1232072|dbj|D11346.1|CHLCP1/1-1545
圖3-15 MN41以18S12放大之定序光譜(反向)(續)
圖3-16 BG33以16S12放大序列演化樹狀圖(Non-protein coding)
圖3-17 BG33以CS12放大序列演化樹狀圖(Non-protein coding)
gi|12547|emb|X65689.1|CSSSRRNA/1-1493
圖3-18 BG33以CS34放大序列演化樹狀圖(Non-protein coding)
圖3-19 BG33以NS12放大序列演化樹狀圖(Non-protein coding)
圖3-20 MN41以18S12放大序列演化樹狀圖(Non-protein coding)
gi|4868347/1-1441
BG CS12/1-709
gi|1232072|dbj|D11346.1|CHLCP1/1-1545
圖3-21 BG33以16S12放大序列演化樹狀圖(Protein coding)
圖3-22 BG33以CS12放大序列演化樹狀圖(Protein coding)
圖3-23 BG33以CS34放大序列演化樹狀圖(Protein coding)
gi|28883285|gb|AY195964.1|/1-2125
圖3-24 BG33以NS12放大序列演化樹狀圖(Protein coding)
圖3-25 MN41以18S12放大序列演化樹狀圖(Protein coding)