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South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private bag X134, Queens wood 0121, Pretoria, South Africa AI Hassen (PhD)

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Page 1: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

South African Rhizobium Culture Collection

(SARCC),WDCM 968

Agricultural Research Council, Plant Protection Research Institute

Private bag X134, Queens wood 0121, Pretoria, South Africa

AI Hassen (PhD)

Page 2: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

Agricultural Research Council (ARC)

• Principal Agricultural research institution in South Africa.

• Conduct research; derive research & development.

• Technology & development transfer.

• Facilitate/ensure natural resources conservation.

• Alleviate poverty.

Page 3: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

• Since 1960s, ARC-Plant Protection Research Institute

• South African Rhizobium Culture Collection (SARCC)

• Several hundreds of rhizobia (>700 strains)

• Long term projects, field collections, client samples,

international institutions

• Rhizobium, Bradyrhizobium, Sinorhizobium, Mesorhizobium

• Routinely maintained (viability, purity )

• WFCC (http://www.wfcc.info/ccinfo/collection/by_id/968)

The SARCC

Page 4: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

• The Nitrogen fixing bacterial symbionts of legumes

• The most important soil microorganisms in agriculture

• Ability of fixing atmospheric nitrogen (N2)

• Biological Nitrogen Fixation (BNF) by Rhizobia spp.

Flavonoids (Legumes) Nod Factors (Rhizobium) Root nodules Bacteroides

• N2 + 3H2 2NH3 (Cost effective) Nitrogenase

Enzyme

Biological Nitrogen Fixation (BNF)

Page 5: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

Screw cap

Nodules

Cotton wool

Desiccant/

Silica gel

Soybean (Glycine max )

nodules ( V-15 growth stage)

Preservation of

nodules in desicant

SARCC- major step in isolation of rhizobia

Page 6: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

• Imbibe nodules in water 1hr.

• Surface sterilize nodules

• Transfer into 100µl sterile water

• Crush with sterile rod & mix

• Plate/streak 30µl suspension

• Yeast Mannitol Agar (YMA)

Streaking on YMA-CR agar

Isolation of Rhizobia from nodules

Page 7: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

a b c

Bradyrhizobium (a) and Rhizobium (b) species

usually form white or transparent colonies on

YMA-CR medium (do not absorb congo red dye)

Azosprillum absorbs the

congo red dye and forms

red colonies on YMA-CR

• Gram negative, rod shaped ( 1.2 - 3µm )

• Motile (single polar/2-6 peritrichous fla.

• No endospore

Isolation and preliminary characterization

Page 8: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

Isolation and characterization

Page 9: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

Preservation of SRACC cultures

Frozen 20% Glycerol Lyophilization

Page 10: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

• Fix atmospheric N, promote growth & yield

• Source of protein for humans and grazing animals

• Improve soil fertility through BNF

• Rhizobia for soybeans, cowpea, beans & chickpea

• Rhizobia for Indigofera spicata var. spicata, Macrotyloma

axillarae, Vigna unguiculata (Indigenous to South Africa)

• Stylosanthus gracilis, Desmodium uncinatum, Indigofera

hirusta, Cyamopsis tetragonoloba, Onchobrychis vicifolia

(exotic species)

Rhizobium holdings at SARCC

Page 11: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

• Glasshouse Koch’s expt. & field trails

• Leonard jar (Hoagland's soln.)

Screening for Effective Rhizobium spp.

1 2

Page 12: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

TJ14

SR4

SAC2

Rhizobium rhizogenes 163C

UD5

Rhizobium tropici strain LNW10

Rhizobium tropici strain LNP6

XCR6

RAK1

Sinorhizobium meliloti QTX 8-2

Sinorhizobium meliloti Qtx-8-1

RF14

Rhizobium taeanense strain PSB 2-6

XAP2

Mesorhizobium amorphae

XEL1

Mesorhizobium sp. SOD33

Azospirillum brasilense REC3

Azospirillum brasilense strain 13-2C

Bradyrhizobium elkanii DASA35013

XHX1

WB74

Bradyrhizobium japonicum USDA 110

Bradyrhizobium japonicum DASA38025

XHT1

XBQ5

Sinorhizobium fredii USDA 194

Mesorhizobium loti

Bradyrhizobium sp. USDA 4424

Bradyrhizobium sp. AP28 FN566882.1 89

68

98

72

58

56

54

100

90

79

99

98

60

94

99 91 90

81 89

98

78

94

99

0.1

• fD1(5’AGAGTTTGATCCTGGCTCAG3)

• rD1(5’AAGGAGGTGATCCAGCC3’)

• Sequencing

• Bioedit, Chromas Lite

• MAFFT

• MEGA5

M a b c d e f g h i j k l m n o p q r s t u nc

16S rRNA

Identification of Effective Strains

Page 13: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

B. japonicum strain SEMIA 6439

SA59-2A

XBD2

Bradyrhizobium sp. CCBAU 05327

XBV7

XFH3

SA59-2F

XBT2

B. liaoningen se CCBAU 13045

Bradyrhizobium genosp.TUXTLAS-16

XHG1

SA59-2E

B. japonicum CCBAU 53152

XHF12

XHF14

XBU4

XCG2

XGE1

Bradyrhizobium elkanii (AB231916.1)

XGA1

XBQ5

SA59-2C

XGO1

XHH1

Rhizobium sp. Qtx 14 2

Sinorhizobium meliloti (FM178848.1)

XFH1

Rhizobium sp. CCBAU 15278(GU552897.1)

Rhizobium etli CFN 42 (U28916.1)

Rhizobium leguminosarum WSM1325

Rhizobium tropici (GQ287633.1)

Methylobacterium cerastii (FR733885.1)

Methylobacterium nodulans ORS2060

100

64 54

91

58

50

10

100

67

99

100

78 86

76

67

68 56

81

61

0.01

- Onchobrychis vicifolia

- Vigna sp.

- Indigofera hirusta

- Cyamopsis tetragonoloba

- Vigna unguiculata

- Macrotyloma axilarae

- Stylosanthus gracilis

- Desmodium uncinatum

Identification of Effective Strains

Page 14: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

Glycine max

(Soybean)

10 20 30 40 50....|....| ....|....| ....|....| ....|....| ....|....|

WB74 ATGCAAGTCG AGCGGGCGTA GCAATACGTC AGCGGCAGAC GGGTGAGTAA 50

B.japonicum strain CCBAU -------TCG AGCGGGCGTA GCAATACGTC AGCGGCAGAC GGGTGAGTAA 43

60 70 80 90 100....|....| ....|....| ....|....| ....|....| ....|....|

WB74 CGCGTGCGAA CGTACCTTTT GGTTCGGAAC AACACAGGGA AACTTGTGCT 100

B.japonicum strain CCBAU CGCGTGGGAA CGTACCTTTT GGTTCGGAAC AACACAGGGA AACTTGTGCT 93

110 120 130 140 150....|....| ....|....| ....|....| ....|....| ....|....|

WB74 AATACCGGAT AAGCCCTTAC GGGGAAAGAT TTATCGCCGA AAGATCGGCC 150

B.japonicum strain CCBAU AATACCGGAT AAGCCCTTAC GGGGAAAGAT TTATCGCCGA AAGATCGGCC 143

160 170 180 190 200....|....| ....|....| ....|....| ....|....| ....|....|

WB74 CGCGTCTGAT TAGCTAGTTG GTAGGGTAAC GGCCTACCAA GGCGACGATC 200

B.japonicum strain CCBAU CGCGTCTGAT TAGCTAGTTG GTAGGGTAAC GGCCTACCAA GGCGACGATC 193

210 220 230 240 250....|....| ....|....| ....|....| ....|....| ....|....|

WB74 AGTAGCTGGT CTGAGAGGAT GATCAGCCAC ATTGGGACTG AGACACGGCC 250

B.japonicum strain CCBAU AGTAGCTGGT CTGAGAGGAT GATCAGCCAC ATTGGGACTG AGACACGGCC 243

260 270 280 290 300....|....| ....|....| ....|....| ....|....| ....|....|

WB74 CAAACTCCTA CGGGAGGCAG CAGTGGGGAA TATTGGACAA TGGGGGCAAC 300

B.japonicum strain CCBAU CAAACTCCTA CGGGAGGCAG CAGTGGGGAA TATTGGACAA TGGGGGCAAC 293

310 320 330 340 350....|....| ....|....| ....|....| ....|....| ....|....|

WB74 CCTGATCCAG CCATGCCGCG TGAGTGATGA AGGCCCTAGG GTTGTAAAGC 350

B.japonicum strain CCBAU CCTGATCCAG CCATGCCGCG TGAGTGATGA AGGCCCTAGG GTTGTAAAGC 343

360 370 380 390 400....|....| ....|....| ....|....| ....|....| ....|....|

WB74 TCTTTTGTGC GGGAAGATAA TGACGGTACC GCAAGAATAA GCCCCGGCTA 400

B.japonicum strain CCBAU TCTTTTGTGC GGGAAGATAA TGACGGTACC GCAAGAATAA GCCCCGGCTA 393

410 420 430 440 450....|....| ....|....| ....|....| ....|....| ....|....|

WB74 ACTTCGTGCC AGCAGCCGCG GTAATACGAA GGGGGCTAGC GTTGCTCGGA 450

B.japonicum strain CCBAU ACTTCGTGCC AGCAGCCGCG GTAATACGAA GGGGGCTAGC GTTGCTCGGA 443

460 470 480 490 500....|....| ....|....| ....|....| ....|....| ....|....|

WB74 ATCACTGGGC GTAAAGGGTG CGTAGGCGGG TCTTTAAGTC AGGGGTGAAA 500

B.japonicum strain CCBAU ATCACTGGGC GTAAAGGGTG CGTAGGCGGG TCTTTAAGTC AGGGGTGAAA 493

510 520 530 540 550....|....| ....|....| ....|....| ....|....| ....|....|

WB74 TCCTGGAGCT CAACTCCAGA ACTGCCTTTG ATACTGAAGA TCTTGAGTTC 550

B.japonicum strain CCBAU TCCTGGAGCT CAACTCCAGA ACTGCCTTTG ATACTGAAGA TCTTGAGTTC 543

560 570 580 590 600....|....| ....|....| ....|....| ....|....| ....|....|

WB74 GGGAGAGGTG AGTGGAACTG CGAGTGTAGA GGTGAAATTC GTAGATATTC 600

B.japonicum strain CCBAU GGGAGAGGTG AGTGGAACTG CGAGTGTAGA GGTGAAATTC GTAGATATTC 593

610 620 630 640 650....|....| ....|....| ....|....| ....|....| ....|....|

WB74 GCAAGAACAC CAGTGGCGAA GGCGGCTCAC TGGCCCGATA CTGACGCTGA 650

B.japonicum strain CCBAU GCAAGAACAC CAGTGGCGAA GGCGGCTCAC TGGCCCGATA CTGACGCTGA 643

660 670 680 690 700....|....| ....|....| ....|....| ....|....| ....|....|

WB74 GGCACGAAAG CGTGGGGAGC AAACAGGATT AGATACCCTG GTAGTCCACG 700

B.japonicum strain CCBAU GGCACGAAAG CGTGGGGAGC AAACAGGATT AGATACCCTG GTAGTCCACG 693

710 720 730 740 750....|....| ....|....| ....|....| ....|....| ....|....|

WB74 CCGTAAACGA TGAATGCCAG CCGTTAGTGG GTTTACTCAC TAGTGGCGCA 750

B.japonicum strain CCBAU CCGTAAACGA TGAATGCCAG CCGTTAGTGG GTTTACTCAC TAGTGGCGCA 743

Glycine max: SARCC-WB74 = 100% Bradyrhizobium spp.

Page 15: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

10 20 30 40 50....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 ---------- ---------- ---------- ---------- -------KAY 3

R.leguminosarum strain CIAM AACGAACGCT GGCGGCAGGC TTAACACATG CAAGTCGAGC GCCCCGC--- 47

60 70 80 90 100....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 AARGGGAGCG GCAGACGGGT GAGTAACGCG TGGGAATCYA CCCTTGACTA 53

R.leguminosarum strain CIAM AAGGGGAGCG GCAGACGGGT GAGTAACGCG TGGGAATCTA CCCTTGACTA 97

110 120 130 140 150....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 CGGAATAACG CAGGGAAACT TGTGCTAATA CCGTATGTGT CCTTCGGGAG 103

R.leguminosarum strain CIAM CGGAATAACG CAGGGAAACT TGTGCTAATA CCGTATGTGT CCTTCGGGAG 147

160 170 180 190 200....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 AAAGATTTAT CGGTCAAGGA TGAGCCCGCG TTGGATTAGC TAGTTGGTGG 153

R.leguminosarum strain CIAM AAAGATTTAT CGGTCAAGGA TGAGCCCGCG TTGGATTAGC TAGTTGGTGG 197

210 220 230 240 250....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 GGTRAAGKCS TAMCARGGMR ACGATCCATA GCTGGTCTGA GAGGATGATC 203

R.leguminosarum strain CIAM GGTAAAGGCC TACCAAGGCG ACGATCCATA GCTGGTCTGA GAGGATGATC 247

260 270 280 290 300....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 AGCCACATTG GGACTGAGAC ACGGCCCAAA CTCCTACGGG AGGCAGCAGT 253

R.leguminosarum strain CIAM AGCCACATTG GGACTGAGAC ACGGCCCAAA CTCCTACGGG AGGCAGCAGT 297

310 320 330 340 350....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 GGGGAATATT GGACAATGGG CGCAAGCCTG ATCCAGCCAT GCCGCGTGAG 303

R.leguminosarum strain CIAM GGGGAATATT GGACAATGGG CGCAAGCCTG ATCCAGCCAT GCCGCGTGAG 347

360 370 380 390 400....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 TGATGAAGGC CCTAGGGTTG TAAAGCTCTT TCACCGGAGA AGATAATGAC 353

R.leguminosarum strain CIAM TGATGAAGGC CCTAGGGTTG TAAAGCTCTT TCACCGGAGA AGATAATGAC 397

410 420 430 440 450....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 GGTATCCGGA GAAGAAGCCC CGGCTAACTT CGTGCCAGCA GCCGCGGTAA 403

R.leguminosarum strain CIAM GGTATCCGGA GAAGAAGCCC CGGCTAACTT CGTGCCAGCA GCCGCGGTAA 447

460 470 480 490 500....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 TACGAAGGGG GCTAGCGTTG TTCGGAATTA CTGGGCGTAA AGCGCACGTA 453

R.leguminosarum strain CIAM TACGAAGGGG GCTAGCGTTG TTCGGAATTA CTGGGCGTAA AGCGCACGTA 497

510 520 530 540 550....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 GGCGGATCGA TCAGTCAGGG GTGAAATCCC AGGGCTCAAC CCTGGAACTG 503

R.leguminosarum strain CIAM GGCGGATCGA TCAGTCAGGG GTGAAATCCC AGGGCTCAAC CCTGGAACTG 547

560 570 580 590 600....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 CCTTTGATAC TGTCGATCTG GAGTATGGAA GAGGTGAGTG GAATTCCGAG 553

R.leguminosarum strain CIAM CCTTTGATAC TGTCGATCTG GAGTATGGAA GAGGTGAGTG GAATTCCGAG 597

610 620 630 640 650....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 TGTAGAGGTG AAATTCGTAG ATATTCGGAG GAACACCAGT GGCGAAGGCG 603

R.leguminosarum strain CIAM TGTAGAGGTG AAATTCGTAG ATATTCGGAG GAACACCAGT GGCGAAGGCG 647

660 670 680 690 700....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 GCTCACTGGT CCATTACTGA CGCTGAGGTG CGAAAGCGTG GGGAGCAAAC 653

R.leguminosarum strain CIAM GCTCACTGGT CCATTACTGA CGCTGAGGTG CGAAAGCGTG GGGAGCAAAC 697

710 720 730 740 750....|....| ....|....| ....|....| ....|....| ....|....|

SAC2 AGGATTAGAT ACCCTGGTAG TCCACGCCGT AAACGATGAA TGTTAGCCGT 703

R.leguminosarum strain CIAM AGGATTAGAT ACCCTGGTAG TCCACGCCGT AAACGATGAA TGTTAGCCGT 747

Trifolium vesiculosum

(Woogenlup clover)

Trifolium sp: SARCC-SAC2 = 100% Bradyrhizobium spp.

Page 16: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

10 20 30 40 50....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 ---------- ---------- ---------- ---------- ---------- 1

R.leguminosarum bv. viciae 384 AGAGTTTGAT CCTGGCTCAG AACGAACGCT GGCGGCAGGC TTAACACATG 50

60 70 80 90 100....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 ---------- ---------- ---------- ---------- -------tgg 3

R.leguminosarum bv. viciae 384 CAAGTCGAGC GCCCCGCAAG GGGAGCGGCA GACGGGTGAG TAACGCGTGG 100

110 120 130 140 150....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 gaatctaccc ttgactacgg aataacgcag ggaaacttgt gctaataccg 53

R.leguminosarum bv. viciae 384 GAATCTACCC TTGACTACGG AATAACGCAG GGAAACTTGT GCTAATACCG 150

160 170 180 190 200....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 tatgtgtcct tcgggagaaa gatttatcgg tcaaggatga gcccgcgttg 103

R.leguminosarum bv. viciae 384 TATGTGTCCT TCGGGAGAAA GATTTATCGG TCAAGGATGA GCCCGCGTTG 200

210 220 230 240 250....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 gattagctag ttggtggggt aaaggcctac caaggcgacg atccatagct 153

R.leguminosarum bv. viciae 384 GATTAGCTAG TTGGTGGGGT AAAGGCCTAC CAAGGCGACG ATCCATAGCT 250

260 270 280 290 300....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 ggtctgagag gatgatcagc cacattggga ctgagacacg gcccaaactc 203

R.leguminosarum bv. viciae 384 GGTCTGAGAG GATGATCAGC CACATTGGGA CTGAGACACG GCCCAAACTC 300

310 320 330 340 350....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 ctacgggagg cagcagtggg gaatattgga caatgggcgc aagcctgatc 253

R.leguminosarum bv. viciae 384 CTACGGGAGG CAGCAGTGGG GAATATTGGA CAATGGGCGC AAGCCTGATC 350

360 370 380 390 400....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 cagccatgcc gcgtgagtga tgaaggccct agggttgtaa agctctttca 303

R.leguminosarum bv. viciae 384 CAGCCATGCC GCGTGAGTGA TGAAGGCCCT AGGGTTGTAA AGCTCTTTCA 400

410 420 430 440 450....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 ccggagaaga taatgacggt atccggagaa gaagccccgg ctaacttcgt 353

R.leguminosarum bv. viciae 384 CCGGAGAAGA TAATGACGGT ATCCGGAGAA GAAGCCCCGG CTAACTTCGT 450

460 470 480 490 500....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 gccagcagcc gcggtaatac gaagggggct agcgttgttc ggaattactg 403

R.leguminosarum bv. viciae 384 GCCAGCAGCC GCGGTAATAC GAAGGGGGCT AGCGTTGTTC GGAATTACTG 500

510 520 530 540 550....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 ggcgtaaagc gcacgtaggc ggatcgatca gtcaggggtg aaatcccagg 453

R.leguminosarum bv. viciae 384 GGCGTAAAGC GCACGTAGGC GGATCGATCA GTCAGGGGTG AAATCCCAGG 550

560 570 580 590 600....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 gctcaaccct ggaactgcct ttgatactgt cgatctggag tatggaagag 503

R.leguminosarum bv. viciae 384 GCTCAACCCT GGAACTGCCT TTGATACTGT CGATCTGGAG TATGGAAGAG 600

610 620 630 640 650....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 gtgagtggaa ttccgagtgt agaggtgaaa ttcgtagata ttcggaggaa 553

R.leguminosarum bv. viciae 384 GTGAGTGGAA TTCCGAGTGT AGAGGTGAAA TTCGTAGATA TTCGGAGGAA 650

660 670 680 690 700....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 caccagtggc gaaggcggct cactggtcca ttactgacgc tgaggtgcga 603

R.leguminosarum bv. viciae 384 CACCAGTGGC GAAGGCGGCT CACTGGTCCA TTACTGACGC TGAGGTGCGA 700

710 720 730 740 750....|....| ....|....| ....|....| ....|....| ....|....|

TJ14 aagcgtgggg agcaaacagg attagatacc ctggtagtcc acgccgtaaa 653

R.leguminosarum bv. viciae 384 AAGCGTGGGG AGCAAACAGG ATTAGATACC CTGGTAGTCC ACGCCGTAAA 750

Pisum sativum

Peas

Pisum sativum: SARCC-TJ14 = 100% Bradyrhizobium spp.

Page 17: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

SARCC data base – Card System

Page 18: South African Rhizobium Culture Collection …South African Rhizobium Culture Collection (SARCC),WDCM 968 Agricultural Research Council, Plant Protection Research Institute Private

• SARCC – South Africa’s National Assets (>700 strains)

• Commercial strains used elsewhere in the world

• Most of the rhizobial strains actively nodulating/ N2 fixing

• Lack of skilled man power, modern Lab facilities

• Conventional method of identification

• Recent improvement , new information to SARCC

- Research Legume-Rhizobium symbiosis

- Identification (molecular tools)

• Development of the SARCC

Concluding Summary