167

Chng 9
Vi khun quang hp v c nh m

I.Vi sinh vt quang hp

1.Cc vi khun quang quang hp (Phototrophic bacteria)
1.Vi khun lu hunh mu ta (Purple sulfur bacteria):
a-HChromatiaceae:
b-HEctothiorhodospiraceae:
2-Vi khun khng lu hunh mu ta (Nonsulfure purple bacteria)
3.Vi khun lu hunh mu lc (Green sulfure bacteria)
4.Vi khun khng lu hunh mu lc (Green nonsulfur bacteria)
5-Vi khunlam(Ngnh Cyanobacteria)
a- Nhm I (c tc gi gi l b Chroococcales):
b- Nhm II (c tc gi gi l b Pleurocapsales):
c- Nhm III (c tc gi gi l b Oscillatorriales):
d- Nhm IV (c tc gi gi l b Nostocales) :
e- Nhm V (c tc gi gi l b Stigonematales)
1.1.Vi khun lu hunh mu ta (Purple sulfur bacteria)
Thuc nhm ny l cc vi khun k kh bt buc, c kh nng quang
t dng v c (photolithoautotroph), t bo c cha chlorophyll a hoc b
, h thng quang hp cha cc mng hnh cu hay hnh phin (lamellar)
gn vi mng sinh cht. dng lm ngun cho in t (electron donors)
trong quang hp thng s dng H2, H2S hay S . C kh nng di ng vi
tin mao mc cc, c loi chu mao, t l G+C l 45-70%.
a- H Chromatiaceae:
1.1.Chi Thiospirium
1.2. Chi Chromatium
1.3. Chi Thiocapsa
1.4. Chi Thiocystis
1.5. Chi Thiospirillum
1.6. Chi Thiorhodovibrio
1.7. Chi Amoebobacter
1.8. Chi Lamprobacter
1.9. Chi Lamprocystis
1.10.Chi Thiodyction
 168

1.11.Chi Thiopedia
1.12. Chi Rhabdochromatium
1.13. Chi Thiorhodococcus

Chromatium Thiocapsa

Thiocystis Thiospirillum

Lamprocystis Thiopedia
Hnh 9.1: Mt s i din vi khun lu hunh mu ta thuc H
Chromatiaceae
 169

b- H Ectothiorhodospiraceae:
1.1- Chi Ectothiorhodospirace
1.2- Chi Halorhodospira
1.2-Vi khun khng lu hunh mu ta (Nonsulfure purple bacteria)
Vi khun khng lu hunh mu ta l nhm vi khun quang d
dng hu c (photoorganoheterotrophs) thng k kh bt buc, mt s
loi l quang t dng v c khng bt buc (trong ti l ho d dng
hu c- chemoorganoheterotrophs). T bo cha chlorophyl a hoc b, h
thng quang hp cha cc mng hnh cu hay hnh phin (lamellar) gn
vi mng sinh cht. dng lm ngun cho in t (electron donors)
trong quang hp thng s dng cht hu c, i khi s dng hp cht
lu hunh dng kh hoc H2. C kh nng di ng vi tin mao mc
cc, hoc khng di ng, mt s loi c ti kh (gas vesicles), t l G+C l
61-72%.
2.1- Chi Blastochloris
2.2- Chi Phaeospirillum
2.3- Chi Rhodobacter
2.4- Chi Rhodobium
2.5- Chi Rhodocista
2.6- Chi Rhodocyclus
2.7- Chi Rhooferax
2.8- Chi Rhodomicrobium
2.9- Chi Rhodoplanes
2.10-Chi Rhodopila
2.11- Chi Rhodopseudomonas
2.12- Chi Rhodospira
2.13- Chi Rhodospirillum
2.14- Chi Rhodothalassium
2.15- Chi Rhodovibrio
2.16-Chi Rhodovulum
2.17- Chi Rosespira
2.18- Chi Rubiviva
 170

Rhodospirillum Rhodospirillum di KHV in t

Rhodopseudomonas Rhodopseudomonas di KHV in t

Rhodobacter Rhodopila

Rhodocyclus purpureus Rhomicrobium

Hnh 9.2: Mt s i din vi khun khng lu hunh mu ta
 171

1.3.Vi khun lu hunh mu lc (Green sulfure bacteria)

Thuc nhm ny l cc vi khun k kh bt buc, c kh nng quang
t dng v c (photolithoautotroph), t bo c cha chlorophyll a cng
vi b , c hoc e, cha caroten nhm 5, h thng quang hp lin quan n
cc lc th (chlorosom) v c lp i vi mng sinh cht. dng lm
ngun cho in t (electron donors) trong quang hp thng s dng H2,
H2S hay S . Ht lu hunh tch lu bn ngoi t bo Khng c kh nng
di ng , mt s loi c ti kh; t l G+C l 48-58%.
3.1- Chi Chlorobium
3.2- Chi Prosthecochloris
3.3- Chi Pelodictyon
3.4- Chi Ancalichliris
3.5- Chi Chloroherpeton

Chlorobium Pelodictyon Prosthecochloris

Hnh 9.3: Mt s i din vi khun lu hunh mu lc

1.4.Vi khun khng lu hunh mu lc (Green nonsulfur bacteria)

Thuc nhm ny l cc vi khun a bo, dng si,thng k kh
khng bt buc ,thng l quang d dng (photoheterotroph), c loi
quang t dng hoc ho d dng. T bo c cha chlorophyll a v c,
trong iu kin k kh thy c chlorosom. dng lm ngun cho in t
(electron donors) trong quang d dng l glucose, axit amin, axit hu c;
trong quang t dng l H2, H2S. Di ng bng phng thc trn
(gliding) , t l G+C l 53-55%. Chi in hnh l Chloroflexus.,
Chloronema
 172

Chloronema Chloroflexus
Hnh 9.4: Mt s i din vi khun khng lu hunh mu lc

1.5- Vi khun lam (Ngnh Cyanobacteria)

Theo NCBT (2005) th Vi khun lam bao gm nhng b sau y:

-Chlorococcales
-Gloeobacteria
-Nostocales
-Oscillatoriales
-Pleurocapsales
-Prochlorales

Trc y thng nhm ln l To lam (Cyanophyta). Thc ra y

l nhng c th nhn nguyn thu, khng lin quan g n to , ngoi kh
nng quang hp hiu kh (quang t dng v c) v dng H2O lm cht
cho in t trong qu trnh quang hp. Vi khun lam cha chlorophyll a
v phycocyanin- phycobiliprotein. Mt s loi c sc t phycoerythrin.
Chng phi hp vi sc t lc to nn mu nu. Mng lin kt vi
phycobilisom. n bo hoc a bo dng si. Khng di ng hoc di ng
bng cch trn (gliding), mt s loi c ti kh (gas vesicles).Nhiu loi
c d t bo (heterocysts) v c kh nng c nh nit. Vi khun lam c
mt khp mi ni, trong t, trn , trong sui nc nng, trong nc
ngt v nc mn. Chng c nng lc chng chu cao hn so vi thc vt
i vi cc iu kin bt li nh nhit cao, pH thp. Mt s loi c kh
nng sng cng sinh vi cc c th khc nh Ru, Dng x, Tu...Nhiu
loi cng sinh vi nm to ra a y (Lichen). Vi khun lam c th l
sinh vt xut hin sm nht trn Tri t
 173

Vi khun lam c chia thnh 5 nhm (subsection) nh sau:

a- Nhm I (c tc gi gi l b Chroococcales):
Hnh que hoc hnh cu n bo, khng c dng si hay dng kt
khi (aggregate); phn i hoc ny chi; khng c d t bo
(heterocytes). Hu ht khng di ng. T l G+C l 31-71% . Cc chi tiu
biu l:
-Chamaesiphon
-Chroococcus
-Gloeothece
-Gleocapsa
-Prochloron

Chamaesiphon Chroococcus

Glooeothece Gleocapsa Prochloron

Hnh 9.5: Mt s i din vi khun lam thuc b Chroococcales
b-Nhm II (c tc gi gi l b Pleurocapsales):
Hnh que hoc hnh cu n bo. c th to dng kt khi
(aggregate); phn ct nhiu ln to ra cc baeocytes; khng c d t
bo.Ch c cc baeocytes l c di ng. T l G+C l 40-46% . Cc chi
tiu biu l:
-Pleurocapsa
-Dermocapsa
-Chroococcidiopsis
 174

Pleurocapsa Dermocapsa Chroococcidiopsis

Hnh 9.6: Mt s i din vi khun lam thuc b Pleurocapsales
c-Nhm III (c tc gi gi l b Oscillatorriales):
Dng si (filamentous) ; dng lng (trichome) khng phn nhnh ch
c cc t bo dinh dng; phn i trn mt phng, c kiu t on
(fragmentation); khng c d t bo; thng di ng. T l G+C l 34-
67%. Cc chi tiu biu l:
-Lyngbya
-Osscillatoria
-Prochlorothrix
-Spirulina
-Pseudanabaena

Lyngbya Oscillatoria Prochlorothrix

Spirulina Pseudanabaena
Hnh 9.7: Mt s i din vi khun lam thuc b Oscillatorriales
 175

d-Nhm IV (c tc gi gi l b Nostocales)
Dng si ; dng lng (trichome) khng phn nhnh c th cha cc
t bo bit ho (specialized cell) ; phn i trn mt phng, c kiu t
on to thnh on sinh sn (hormogonia) ; c t bo d hnh ; thng di
ng c th sn sinh bo t mng dy (akinetes). T l G+C l 38-47%.
Cc chi tiu biu l :
-Anabaena
-Cylindrospermum
-Aphanizomenon
-Nostoc
-Scytonema
-Calothrix

Anabaena Anabaena trong Bo hoa du

Cylindrospermum Calothrix
 176

Nostoc Scytonema
Hnh 9.8: Mt s i din vi khun lam thuc b Nostocales
e-Nhm V (c tc gi gi l b Stigonematales) :
Lng (trichome) dng si, phn nhnh hoc do cc t bo nhiu hn
mt chui to thnh ; phn i theo nhiu mt phng, hnh thnh on sinh
sn (hormogonia) ; c t bo d hnh ; c th sn sinh bo t mng dy (
alkinetes), c hnh thi phc tp v bit ha (differentiation). T l G+C l
42-44%. Cc chi tiu biu l :
-Fischerella
-Stigonema
-Geitlerinema

Stigonema Fischerella Geitlerinema

Hnh 9.9: Mt s i din vi khun lam thuc b Stigonematales

2.Trao i cht cc vi sinh vt quang dng
Tt c cc vi khun quang hp u cha sc t quang hp. Sc t
quang hp vi khun c gi l bacteriochlorophyll. Chlorophyll v
bacteriochlorophyll cn c gi l cht dip lc v cht khun lc. Cht
dip lc, cht khun lc v huyt sc t c cu trc tng t nh nhau.
l mt vng pocphiril do 4 nhn pirol lin kt vi nhau. Li ca cht dip
lc v cht khun lc l Mg, cn li ca huyt sc t l Fe, cht dip lc a
 177

khc vi cht khun lc a,b,c,d,e 7 gc R (t R1 n R7) c th thy r

s khc nhau ny trong bng sau y:
Bng 9.1: S sai khc gia cht dip lc a v cc loi cht khun lc

Ch thch: a gia C3 v C4 khng c ni i, khng thm H

b gia C3 v C4 khng c ni i, c thm H
C th thy r hn s phn bit ca hai loi chlorophyll,
bacteriochlorophyll v hem...bng cch quan st hnh sau y:
 178

H. 9.10: Cu trc ha hc ca mt s sc t quang hp

 179

Ngoi cc loi chlorophyll vi khun t dng quang nng cn c

cha mt s cc sc t thuc loi carotenoit. Carotenoit vi khun khng
ging vi carotenoit to hoc thc vt. Di y l vi v d:

H. 9.11: Cu trc ha hc cc loi carotenoit

vi khun t dng quang nng c hai loi phosphoryl ha quang
hp: phosphoryl ha quang hp tun hon v phosphoryl ha quang hp
khng tun hon.

Ch thch
Bchl*: trng thi kch pht ca
bacteriochlorophyll
Ru-5-P: Ribulose-5-phosphate
H2A: Cht v c cho hydrogen

H.9.12:Phosphoryl ha quang hp kiu tun hon

Trong iu kin k kh mt s vi khun quang hp c th s dng
nng lng ca nh sng thc hin phn ng phosphoryl ha sn sinh
ra ATP. Electron t bacteriochlorophyll c tch ra di tc dng ca
nh sng, sau tham gia vo chui h hp tun hon v quay tr li
Bchl. Trn ng i sn sinh ra ATP. Vic sinh ra ATP c thc hin
ring r vi vic sinh ra [H] c nng lc kh. [H] c nng lc kh c
sinh ra t cc cht v c cho hydrogen (nh H2S...).Qu trnh quang hp
ny khng sn sinh ra oxi.
 180

H.9.12: Phosphoryl ha quang hp kiu khng tun hon

Ch thch: Chlb* v Chla* l trng thi kch pht.

y l kiu phn ng phosphoryl ha quang hp gp vi khun
lam v cng tng t nh to v cy xanh. y chui vn chuyn
electron l khng tun hon v vy cn gi l dng chy electron khng
tun hon.
Quang hp xy ra trong iu kin c oxi v c 2 h thng quang.
H thng quang I c cha Chl a (cht dip lc a) v c th s dng
c tia c bc sng di.
H thng quang II c cha Chl.b v c th s dng c tia sng
lam (bc sng ngn). Qu trnh quang hp ny c sn sinh oxi v xy ra
ng thi vic sinh ATP( h thng quang hp II) v vic sinh [H] c
nng lc kh (trong NAD PH2) l H+ v e-, c sinh ra sau s quang gii
nc.
Sau quang gii nc sn sinh ra O2 +2H+ + 2e, electron s lin
tip i qua chui electron (I v II), sau cng em electron n cho NADP+
sn sinh ra 2 phn ng phophoryl ha sinh ATP. h thng I c
sinh NADPH2 v ATP, cn h thng II c sinh oxi v ATP.
Mt s vi khun a mn t dng quang nng c qu trnh quang
hp kh dc bit. i din cho nhm ny l cc vi khun hay gp trn cc
hi sn p mui (nh vi khun Halobacterium halobium, H.eutirubrum...)
Mng cht t bo ca chng phn thnh 2 phn: phn mu v
phn mu ta. Phn mu c cha cytocrom, flavoprotein.. ca chui h
hp dng oxi ha phosphoryl ha. Phn mu ta rt k l. Trn mng
hin ln cc vt khoang (ng knh khong 0,5m), phn b c lp vi
 181

nhau, chim ti mt na tng din tch ca mng t bo vi khun. Mng

ny c kh nng thc hin mt qu trnh quang hp c o. C mt loi
protein gi l bacteriorodopsin, rt ging vi rodopsin, loi protein c
trong t bo hnh tr trn vng mc ca mt. Bacteriorodopsin chim ti
75% mng mu ta.
Nhng vi khun ny c th sinh trng c trong ti hoc ngoi
sng nu c mt oxi, khi khng c mt oxi chng ch sinh trng c
ngoi sng. Chng c 2 con ng thu nhn nng lng: mt ng oxi
ha phosphoryl ha khi c oxi v mt ng oxi ha phosphoryl ha khi
c chiu sng (quang hp). Tc to ra ATP l cao nht khi c
chiu sng bng bc sng 550-600nm.

H.9.13: S minh ha 2 h thng quang

3. C ch quang hp
3.1. Pha sng quang hp
Pha sng l giai on u ca quang hp, giai on ny nh sng l
nhn t trc tip tham gia vo quang hp nn gi l pha sng.
Pha sng xy ra qua 2 giai on: giai on quang l v giai on
quang ho.
* Giai on quang l: Nh tnh cht quang ho ca nh sng v kh
nng hp th nng lng nh sng ca phn t chlorophyll m nng lng
ca nh sng c chuyn sang nng lng ca in t trong cc phn t
chlorophyll. Nng lng ny li chuyn n 2 tm quang hp l 700 v
680 thc hin giai on quang ho tip.
* Giai on quang ho: quang ho l giai on chuyn ho nng
lng ca cc in t trong 2 tm quang hp c lm giu bi nng
lng nh sng thnh nng lng cha ng trong cc hp cht giu nng
lng l ATP v NADPH2.
Giai on quang ho xy ra ti 2 tm quang hp bi 2 phn ng
 182

quang ho m phn chnh l quang phn ly nc v phosphoryl ho.

- Quang phn ly nc: Nh nng lng nh sng vi s tham gia
ca sc t v h thng cc cht oxi ho trong lc lp m nc b phn hu.
2H2O ( 4H+ + 4 + O2 )
- Phosphoryl ho : y l qu trnh tng hp ATP nh nng lng
ca qu trnh oxi ho xy ra trong chui vn chuyn quang hp.
ADP + H3PO4 ATP + H2O
C 3 hnh thc Phosphoryl ho xy ra:
+ Phosphoryl ho vng.
+ Phosphoryl ho khng vng.
+ Phosphoryl ho ho vng gi.
Kt qu chung ca pha sng l:

12H2O +18ADP +18H3PO4+12NADP+ 18ATP + 12NADPH+H+ + 6O2

+ 18H2O
Pha sng to ra ATP v NADPH+H+ cung cp cho pha ti quang
hp.
3.2. Pha ti quang hp
Sau khi pha sng to ATP v NADPH+H+, giai on tip theo ca
quang hp l s dng ATP v NADPH+H+ kh CO2 to sn phm s
cp ca quang hp l C6H12O6. Qu trnh ny khng cn nh sng nn gi
l pha ti. Pha ti din ra nhiu con ng khc nhau, mi con ng c
trng cho mt nhm thc vt. C 3 con ng ng ho CO2 trong quang
hp: chu trnh Calvin, chu trnh Hatch-Slack v chu trnh CAM.

* Chu trnh Calvin (chu trnh C3)

- y l con ng ng ho CO2 ph bin nht do Calvin tm ra.
Chu trnh xy ra qua 3 giai on chnh:
+ Tip nhn CO2 : Ribulozo 1,5dP tip nhn CO2 sau bin i
thnh 2 phn t APG. APG l sn phm u tin nn chu trnh cn c
gi l chu trnh C3.
+ APG b kh thnh AlPG nh ATP v NADPH+H+ ca pha sng.
+ AlPG s ti to li Ribulozo 1,5dP ng thi to nn sn phm ca chu
trnh l C6H12O6.
Tm tt chu trnh Calvin:
 183

C5 + CO2 + ATP + NADPH ---> C6H12O6

H. 9.14: Chu trnh Calvin

www.starsandseas.com/. ../cellcalvin.htm

II.C inh nitrogen phn t

Trong khng kh c rt nhiu N phn t, nhng tuyt i a s sinh

vt khng s dng c ngun N ny. Ch c mt s VSV l c th hp
th c N. Qua hot ng sng ca chng N s c chuyn thnh N
hp cht (protein v cc sn phm thy phn protein). Hot ng ny c
gi l s c nh N phn t.
Qu trnh c nh N c tc dng rt ln n i sng cc sinh vt
trn tri t. Trn c bn trong nham thch khng cha hp cht N. Cho
nn ngun N trong t hu nh hon ton do tc dng c nh N ca VSV
m hnh thnh. Hng nm cy trng li ly i mt lng N ng k. Hp
cht N c hnh thnh trong qu trnh c nh ny li chnh l ngun N
ch yu b sung cho t.
Cc loi VSV c nh N khng nhiu lm, c th chia lm hai loi
ln nh sau:
1.Vi sinh vt c nh N sng t do (khng cng sinh): bao gm mt s vi
khun, x khun. Trong s ny quan trng nht l cc loi thuc chi
Azotobacter v loi Clostridium Pasteurianum.
1. Azotobacter
1.1.S lc v Azotobacter
Azotobacter l vi khun c nh N sng t do trong t, hiu kh,
khng c bo t. Chng c phn lp v nui cy thun khit t nm
 184

1901 do nh VSV H Lan Beijerinck. i din in hnh ca chi ny l

Azotobacter chroococcum.
Do iu kin nui cy v la tui khc nhau m Azotobacter c hnh
thi thay i. Lc cn non Azotobacter l nhng trc khun mp hnh que,
c tin mao, c kh nng di ng, khi gi th tr thnh t bo hnh cu c
gip mc xung quanh.
Azotobacter c th s dng nhiu loi hp cht hu c lm ngun
thc n C. Chng cng cn nhiu nguyn t khong, c bit l 2 nguyn
t vi lng bor (B) v molipbden (Mo)(Mo cn cho tc dng c nh N).
Khi sng trong iu kin khng c N hp cht Azotobacter s dng
N ca khng kh bin thnh hp cht N ca c th sng. Khi sng trong
mi trng thc n N hu c hoc v c th tc dng c nh N s rt
thp hoc khng c. Azotobacter thch hp vi iu kin hiu kh va phi
v pH trung tnh hoc hi kim. Trong iu kin thch hp c mi khi tiu
th 1gam hydratcarbon Azotobacter c th c nh c 10- 15mg N.
V Azotobacter c kh nng c nh N mnh m nh vy nn t cui
th k 18 c nhiu nc nghin cu s dng chng lm phn vi khun
(gi l phn Azotobacterin). Vic s dng Azotobacterin cng ang pht
trin mnh nhiu nc v thu c kt qu. Tuy nhin s dng
Azotobacterin khng phi l vn tip ging n gin, m cn phi to
iu kin chng pht trin mnh c nh c nhiu N, ngha l s
dng k thut lin hon.
ch to Azotobacterin trc ht ta nui tht nhiu Azotobacter
trn mi trng c hoc lng c bm khng kh. Sau em trn chng
vi than bn hoc cc cht nui dng khc ch thnh bt
Azotobacterin.
1.2. Mt s c im sinh hc ca vi khun Azotobacter
a. Hnh thi khun lc
Khi nui trong mi trng thch,vi khun Azotobacter c khun
lc nhy, li hoc tan, lc u khng mu, sau bin thnh mu nu ti,
thm ch n mu en nhng khng lm nhum mu mi trng khun
lc. Ngoi ra mt s loi Azotobacter c dng nhn nheo, khun lc c
mu vng lc, mu hng .
V nhy ca vi khun Azotobacter cha khong 75% l cht hirit
ca acid uronic v cha khong 0.023% N.
 185

H. 9.15: Hnh dng t bo Azotobacter sp.

(a) di knh hin vi in t (b) tiu bn m
(www-micro.msb.le.ac.uk/ video/Azotobacter.html)

b. Hnh thi t bo vi khun Azotobacter

Azotobacter l loi vi khun Gram m khng sinh bo t. Khi cn
non t bo c dng que u trn ng ring l hay xp thnh tng i
chng cht, cht t bo nhum mu ng u, c kh nng di ng nh
tin mao, kch thc khong :2,0- 7,0 10- 2,5 m . Khi gi t bo
Azotobacter mt kh nng di ng, kch thc thu nh li bin thnh dng
hnh cu. Sinh cht xut hin nhiu ht ln nhn. l cc ht volutin,
granulose, cc git m
Lng ADN trong t bo Azotobacter thng thp hn so vi
nhiu loi vi khun khc (0,70- 0,81%).
Tm li hnh dng t bo Azotobacter v chu k bin i ca chng
ph thuc vo tui ca ng ging v iu kin pht trin.
1.3. Cc c tnh sinh l, sinh ho.
a. c im nui cy :
Vi khun Azotobacter thuc loi vi khun hiu kh, sng theo
phng thc d dng. Chng s dng nhiu loi hp cht hu c khc
nhau: disacarit, dextrin, tinh bt, acid hu c, hp cht thm, tuy vy,
nhiu tc gi cho bit c khng t cc chng Azotobacter khng c kh
nng ng ho lactose, manitol hoc natribenzoat.
b. Kh nng s dng ngun N:
Trn cc mi trng khng cha N khun lc Azotobacter c dng
nhy, li, i khi nhn nheo. Chng t vi khun Azotobacter c kh nng
sinh trng trn mi trng khng c N. S d chng tn ti c l v c
kh nng ng ho mui amonium, ur. Mt s chng Azotobacter c kh
 186

nng s dng nitrit, nitrat. Hai loi acid thch hp nht i vi nhu cu
dinh dng ca Azotobacter l acid glutamic v axtit asparaginic.
1.4. c im phn loi ca vi khun Azotobacter
Theo Becking (1974) th vi khun c nh m thuc chi
Azotobacter c 4 loi: A. chroococcum; A. beijerinckii; A. vinelandii; A.
agilis .
1.5. nh hng cc nhn t sinh thi n s sinh trng v pht trin ca
cc vi khun Azotobacter
Vi khun Azotobacter hiu kh, s pht trin v kh nng c nh
N ca chng trong t chu nh hng ca cc iu kin sau:
a. m ca t
Azotobacter i hi m rt cao ca t. Nhu cu v m ca
chng cao hn so vi cc vi khun khc, tng ng vi nhu cu ca cy
trng. V vy t gp chng vng kh hn, s kh hn ca t ch c bo
xc ca chng mi chu ng c. m thch hp:75- 80%.
b. thong kh .
thong kh ca t c lin quan n qu trnh c nh N ca
Azotobacter. Tuy vy vi khun thuc loi hiu kh nhng c th pht trin
c trong iu kin vi hiu kh. Qu trnh c nh N ca Azotobacter b
gim khi th xi ho kh ca mi trng cao qu +200 mv hoc thp qu
() 200 mv. Nh vy, khng kh qu mnh cng c ch qu trnh c nh
N phn t, khi nng xi trong khng kh l 4% qu trnh c nh N
vt qu ba ln so vi khi xi l 10- 20%.
c . Nhit .
Nhit nh hng n Azotobacter khng ging nhau cc vng
a l khc nhau. vng nhit i Azotobacter thch hp vi nhit 35-
400C. nhit 70C Azotobacter c hot ng c nh N thp hn 5 ln
so vi 450C. T bo sinh dng ca Azotobacter khng sng c khi x
l 500C trong 30 pht, nhit 800C s cht rt nhanh. Nhit thch
hp i vi s pht trin ca Azotobacter vo khong 20-300C.
d. pH ca t.
pH ca t nh hng khng ging nhau n s pht trin v kh
nng c nh N ca Azotobacter. Azotobacter thch hp nht vi pH = 7,2-
8,2, song c th pht trin c pH = 4,5- 9,0. Cc loi Azotobacter
khc nhau mn cm mt cch khc nhau i vi pH ca mi trng.
Chng hn: pH thp nht ca mi trng i vi Azotobacter
 187

chroococcum v A. beijerinck khong 5,5; i vi A. marocytoges l

khong 4,6.
Phn ln Azotobacter ch pht trin pH ln hn 6, v vy t gp
chng t chua. Cng c th phn lp c mt s chng t t chua
nhng cc chng ny thng mt kh nng c nh N phn t.
e. Phn m
Trong t Azotobacter c kh nng ng ho cc cht dinh dng
t mi trng. Ngun N i vi Azotobacter khng phi ch l N phn t
m cn l mui amon, nitrat, amino acid. Tu thuc vo nng ca cc
hp cht cha N c trong mi trng m qu trnh c nh N s b c ch
nhiu hay t .
g. Phn ln.
Pht pho rt cn cho qu trnh c nh N ca Azotobacter ch bt
u xy ra khi nng PO43- t n 4 mg trong 100 ml mi trng.
Ngc li khi nng PO43- t ti 800 mg trong 100 ml qu trnh c nh
N s bt u ngng li. S mn cm mnh m ca Azotobacter vi pht
pho cho php ngi ta s dng chng nh loi vi khun ch th xc nh
nhu cu v phospho ca t.
h. Phn kali.
S pht trin ca Azotobacter rt cn n kali, nhng vi lng
nh. Nu a vo mi trng mt lng mui kali qu d tha chng s
lm c ch s pht trin ca Azotobacter, c th tc hi ny l do gc
anion ca mui ny gy ra.
i. Canxi.
Canxi nh hng ln n s pht trin ca Azotobacter. Khi thiu
canxi t bo Azotobacter s to thnh nhiu khng bo nh hng xu i
vi vic tng hp ATP v s to thnh cc polyphotphat.
Nhng iu tra Vit Nam cho thy trong t c lng cha CaO
cao hn 0,4% lun c mt mt s lng ln cc t bo Azotobacter. Hu
nh khng pht trin trong mu t c c lng cha CaO di 0,25%
(Nguyn Ln Dng 1965)
Ring i vi A.chroococcum nng CaCl2 ,thch hp nht l
0,01% ,nu cao hn s nh hng khng tt i vi hot ng c nh N
ca chng. C tc gi dng A. chroococum nh l loi vi khun ch th
xc nh nhu cu v vi ca t, nguyn t Mg c Azotobacter i
hi vi s lng cao hn st khong 10 ln .
 188

Cng vi nguyn t i lng, cc nguyn t vi lng cng cn

thit i vi Azotobacter, ng ch hn c l molybden, bo(Bo),
mangan(Mn)
k. Cc nhn t sinh hc
S pht trin v c nh N ca Azotobacter trong t cn chu nh
hng mt thit ca khu h cc sinh vt t. Bn cnh cc nhm vi sinh
vt c nh hng tt cn c nhiu nhm c kh nng c ch s pht trin
ca Azotobacter .
c nhiu cng trnh nghin cu cp n mi quan h ga
Azotobacter vi cy trng vi s lng cao hn nhiu so vi ngoi vng
r .
Mt s chng A.chroococum c kh nng sinh ra mt s cht
chng nm c tc dng kh rng (c ch Aspergillus, Penicillium,
Fusarium, Alternaria )
1.6. Tnh hnh nghin cu ng dng Azotobacter.
Trong nhiu nm, nhiu nc khc nhau ,ngi ta sn xut
quy m cng nghip hoc th cng nghip nhng ch phm Azotobacter
v gi l azotobacterin. l nhng dch nui cy Azotobacter c hp
th vo than bn hoc cc loi t giu cht hu c trung ho v b
sung thm mt t phn phospho, kali
Khc vi nitragin, azotobacterin khi dng bn cho cy trng
thng a n nhng hiu qu khng ln v khng n nh (nu c lm
tng sn lng thng cng khng tng qu 10% so vi i chng ).
a s cc tc gi cho rng hiu qu ca vic s dng azotobacterin
ch tng i vi cc t giu cht hu c hoc trong trng hp c
bn thm nhiu phn khong. C l tc dng ch yu ca chng khng
phi kha cnh c nh N m l kha cnh tng hp cc cht hot ng
sinh hc c tc dng kch thch sinh trng ca cy trng .
Vit Nam, vic sn xut v s dng ch phm azotobacterin ch
t ra trong nhng iu kin thm canh c kh nng di do v phn bn,
cn bn vi v bn phn ln lm tng s lng Azotobacter trong t.
Azotobacterin ch c tc dng tng i r i vi cc loi t
giu cht hu c hoc trong trng hp c bn thm nhiu cht
khang.C l tc dng ch yu ca chng khng phi kha cnh c nh
N m l kha cnh tng hp cc cht hot ng sinh hc c tc dng
kch thch s sinh trng ca cy trng .
a. Cc phng php to ch phm Azotobacter
 189

* Dng mi trng thch nghing

Chun b mi trng thch nghing
Thnh phn mi trng (g/l):Glucose-20; K2HPO4.3H2O -0,2;
MgSO4 .7H2O -0,2; NaCl- 0,2; K2SO4 -0,1; CaCO3 -5,0; H20 -1 lt ;Thch -
20 pH: 6,5-7.
Chun b ging Azotobacter :Cc ng ging c phn lp v
tuyn chn t trc.
To ch phm: Cy truyn ging vi khun Azotobacter t ging
gc vo nhng ng nghim thch nghing. m nhit : 28-30 0 C
trong thi gian 2-6 ngy, loi b nhng ng b nhim vi sinh vt khc ta
thu c chng Azotobacter thun khit.
* Dng dch th
Chun b mi trng dch th
Thnh phn mi trng (g/l):
Glucose 20
K2HPO4.3H2O 0,2
MgSO4.7H2O 0,2
NaCl 0,2
K2SO4 0,1
CaCO3 5,0
H2O 1 lt
pH 6,5-7
Chun b ging vi khun Azotobacter :
Ly 5-10 ml mi trng dch th (C thnh phn nh trn khng
c thch) kh trng cho vo mi ng nghim 14 5ml
Dng que cy gt nh, a dch vi khun vo bnh nn (250-
500ml) ng 150ml mi trng v nui chng trn my lc(220 dao
ng/pht) .
Khi khi dch nui t mt 109 t bo/ml th c th em s dng
* Dng bt
Nui cy ging vi khun
Nhn ging
Hp th ging vo cht mang vi khun
im quan trng cua phng php ny l la chn cht mang vi
khun, cht mang vi khun phi t yu cu thch hp v s tn ti ca
Azotobacter trong qu trnh bo qun v sng st trn ht ging khi
c tm vo, cht mang vi khun c th l than bn, t bt giu cht
hu c, c ni s dng c tru, b bia lm cht mang vi khun
 190

Ngy nay cht mang vi khun c s dng bi than bn thch hp

trong vic s dng ch phm vi khun ni chung v ch phm Azotobacter
ni ring. Ngoi c tnh ca than bn l khng c, c tnh hp th cao,
gi nc tt , n cn l nguyn liu r tin sn c nhiu ni.
1.2. Clostridium pasteurianum v mt s VSV c nh N khc
C.pasteurianum l trc khun k kh bt buc, c bo t, bo t nm
gia lm t bo phnh ln nh hnh thoi.
C. pasteurianum l mt loi vi khun butyric, chng c th s dng
cc hydratcarbon thng thng v lm ln men suinh ra acid butyric,
acetic, CO2 v H2. Chng c th s dng hp cht N v c v hu c lm
ngun thc n N, khi nhng hp cht ny y th tc dng c nh N
ca chng h thp hoc hon ton khng c.
Tc dng c nh N ca C. pasteurianum thp hn Azotobacter. C
mi khi tiu th 1g hydratcarbon th chng c nh c 2-3 mg N. Tuy
nhin C.Pasteurianum rt nhiu v phn b rng ri hn Azotobacter nn
ngun N m chng c nh cho t l rt quan trng. Chng li pht trin
c rung ngp nc, yu cu pH khng gt gao nn thch hp cho cc
loi rung ca ta.
Ngoi hai loi trn, trong cc VSV c nh N cn c mt s vi
khun thuc chi Clostridium, Bacillus v Azotomonas. Tuy nhin, v
nhng loi ny phn b khng nhiu v hiu lc c nh N cng thp nn
tc dng thc t khng ln lm.
2. Vi khun c nh N cng sinh.
Phn ln cc cy thuc h u khng cn thc n hp cht, chng c
th s dng c N khng kh. Nhng kh nng ny khng phi bn thn
cy h u c m do nhng vi khun nt r cng sinh vi cy h u
mang li.
2.1. Vi khun nt r
Ngi u tin chng minh rng cc cy b u c th sinh trng
trong t khng cha N l H. Hellrigel v H. Wilfarth (1886).
Nm 1886 M.W.Beijerinck phn lp c vi khun t nt r
mt s cy u.
Ging (chi) Rhizobium (Franck, 1889) l loi vi khun n bo
t, rt d nhm vi Pseudomonas, trc khun t do c kch thc 1,2(m x
0,5 chuyn ng nh nhung mao v tin mao cc, hiu kh. Khi hnh
thnh nt sn th trc khun bin dng (c hin tng a hnh thi), n
thay i hnh dng ty theo la tui v iu kin sng. Khi cn non chng
l trc khun nh b, khng c bo t, di ng nh tin mao. Trong qu
trnh pht trin, chng s chuyn thnh hnh cu v nhng dng phn
nhnh hnh ch Y, ch V gi l gi khun th (Bacteroids).
 191

Nm 1963, Manil chia cc vi khun nt sn thnh 3 nhm:

-Rhizobium leguminosa (u H Lan), R. trifolii (c ba l), R.
phaseoli (u c ve, u xanh).
-Rhizobium lupini (mc tc, linh lng), gp nhm 1 v 2 thnh
Rhizobium mc nhanh.
- Rhizobium lupini (u a, lupin), R. Japonicum (u nnh) vi
nhiu loi ph. Nhm ny hin nay c nh loi l Bradyrhizobium
(gm tt c cc chng mc chm.
Hin nay, ngi ta xp vi khun nt r vo mt chi chung l
Rhizobium.Chng c th s dng nhiu hydratcarbon khc nhau lm
ngun C. Khi sng ring l trong t hoc trong mi trng nui cy
chng khng c kh nng c nh N.
V h hp, chng l loi vi hiu kh. Rhizobium c nhiu loi v c
tnh cht chuyn tnh, ngha l mi loi ch c th xm nhp v to thnh
nt r trong mt s cy h u nht nh.
Vi khun nt r khi gp cy h u thch hp s xm nhp vo t
bo r, kch thch r to thnh nhng nt r. Nt r thng thng bng
ht go, nt r to bng ht u nh, trong nt r cha y vi khun. Sau
khi hnh thnh nt r gia vi khun v cy h u lm thnh mt th cng
sinh c li cho c hai bn.
Ngoi cc cy h u, mt s r cy khc cng c th cng sinh vi
vi khun c nh N. Gn y ngi ta cn thy nt sn l mt vi loi
cy nhit i. Trong nt sn cng cha vi khun c nh N gn ging
r.
2.2.Phn bn nt r (nitragin)
Phn bn vi sinh do Noble Hiltner sn xut u tin ti c nm
1896 v c t tn l nitragin: Sau pht trin sn xut ti mt s
nc khc nh M (1896), Canada (1905), Nga (1907), Anh (1910) v
Thy in (1914).
Nitragin l loi phn c ch to bi vi khun Rhizobium do
Beijerink phn lp nm 1888 v c Fred t tn vo nm 1889 dng
bn cho cc loi cy thch hp ca h u. T cho n nay c nhiu
cng trnh nghin cu nhm ng dng v m rng vic sn xut cc loi
phn bn vi sinh c nh nit m thnh phn cn c phi hp thm mt
s vi sinh vt c ch khc nh mt s x khun c nh nit sng t do
Frankia spp, Azotobacter spp, cc vi khun c nh N sng t do
Clostridium pasteurianum, Beijerinkia indica, cc x khun c kh nng
gii cellulose, hoc mt s chng vi sinh vt c kh nng chuyn ha cc
 192

ngun d tr phospho v kali dng kh ho tan vi s lng ln c

trong t mn, than bn, trong cc qung apatit, phosphorid v.v... chuyn
chng thnh dng d ho tan, cy trng c th hp th c.
Trong cc loi vi sinh vt k trn, vi khun nt r c ngha ht sc
to ln trong nng nghip. Cng c nh N cng sinh ln hn nhiu so
vi cng c nh N khng cng sinh. Sau khi trng cc cy h u t
giu thm N rt nhiu. Chnh v vy, t u th k XX ngi ta nghin
cu s dng phn bn nt r vi mc ch l tip ging nhn to, lm
cho t khng cha vi khun nt r thch ng s c vi khun thch ng,
lm cho cy h u khng sinh nt r s c th sinh nt r v nhng cy
c th sinh nt r th cng sinh nhiu hn, c nh N cng mnh m hn.
Cch s dng phn bn vi khun nt r l em trn phn vi ht
ging, ri kh, sau em gieo. Cng c th dng nt r tt, nghin nt
trn vi ht ging. Khi s dng c th tng sn lng cy h u ln
15- 20%.
Ngoi hai nhm vi khun c nh N sng cng sinh vi thc vt v
sng t do trong t nh ni trn, cn mt s vi khun c kh nng
c nh N sng trn b mt r v n su vo vo lp t chc b mt r ca
mt s cy ha tho nh la, ng, ma,... l mt loi vi khun c dng
xon c pht hin t nm 1974 thuc chi Azospirillum.
Ngoi cc nhm vi khun c nh N ni trn ra, cn c mt s loi
vi to n bo cng c kh nng c nh N. V d nh vi khun lam sng
t do v sng cng sinh trong bo hoa du. Cc loi ny cng ng gp
khng nh vo qu trnh c nh N khng kh.
Cu hi n tp chng 9

1.Hy nu tng qut chu trnh chuyn ha N trong thin nhin v vai tr
ca vi sinh vt trong chu trnh ?
2.Cc nhm vi sinh vt c nh N ? ngha thc t ca vic nghin cu vi
sinh vt c nh m.
3.Nu mt s thnh tu hin nay v nghin cu vi sinh vt c nh m.
4. Naza l g ? C ch hin bit ca qu trnh c nh N phn t.
5.Cc loi phn bn vi sinh vt ? c s khoa hc ca vic sn xut v s
dng cc loi phn bn vi sinh vt?
6. Trnh by cc bc ch yu iu ch Azotobacterin v Nitragin. Trong
iu kin hin nay Vit Nam c th sn xut 2 ch phm ny c
khng ? Ti sao ?