BSCI 442  Lec. # 20-21     N ASSIMILATION: OUTLINE       Slides                   F 2008

ONLY PLANTS AND MICROORGANISMS ARE ABLE TO OBTAIN N FROM BOTH AIR AND SOIL.
N CYCLE

TWO WAYS PLANTS CONVERT AVAILABLE N TO A BIOL. USEFUL FORM
I.  Biological N Fixation: N2 ---.> NH3
II. NO3- Reduction:   NO3- ---> NH4+
NH4+ --> Amides or Ureides --> Amino acids --> Proteins

NITRATE REDUCTION
 ALL PLANTS CAN TAKE UP N IN THE FORM OF NH4+ and NITRATE (NO3-).  NITRATE MUST BE REDUCED TO NH4+ BEFORE IT CAN BE INCORPORATED INTO AMINO ACIDS, PROTEINS & NUCLEIC ACIDS.
a. Nitrate  Reductase
NO3- + NADPH + H+ ---------> NO2- + NADP + H2O
b. Nitrite Reductase
NO2- + 6 Fdred + 8H+----> NH4+ + 2H2O

NH4+ IS ASSIMILATED INTO ORGANIC N BEFORE IT CAN BE EXPORTED.   NH4+ IS INCORPORATED INTO
I. AMIDES (ASP-NH2) or
II. UREIDES
And then TRANSPORTED ELSEWHERE

AMIDES OR UREIDES --> AMINO ACIDS ---> PROTEINS
AMIDES OR UREIDES ---> NUCLEIC ACIDS --> DNA, RNA

TWO TYPES OF BIOL. N FIXATION
1. FREE LIVING N FIXING BACTERIA [4%]. eg.CLOSTRIDIUM
2. SYMBIOTIC N FIXERS [90%]. e.g. LEGUME-RHIZOBIUM

SYMBIOTIC N FIXATION IN LEGUMES INITIATES WHEN N IS LIMITING-
 HOW DOES IT START AND WHAT DOES IT LEAD TO?
A) Recognition and binding : IT’S ALL CHEMISTRY!
B) Nodulation:
    BACTERIA ENTER THE PLANT ROOT THROUGH AN INFECTION THREAD
    CORTICAL CELLS OF ROOT DIVIDE (HORMONE-STIMULATED) TO FORM NODULE
    BACTERIA DIFFERENTIATE INTO BACTERIODS
C) Bacteriods develop ability to fix N gas.

Development is dependent on the regulated expression of bacterial and plant genes.
Plant genes: nodulins
Bacterial genes: Nif, Nod, Fix

NITROGENASE in BACTEROID.  HOW IS ATM. N FIXED?
By one reaction catalyzed by nitrogenase.  .
N2 + 8e- + 8H+ + 16 ATP --> 2NH3 + H2 + 16 ADP + 16 Pi
Requires lots of energy and reducing power. .
Nitrogenase is inactivated by oxygen.
Inactivation is protected by leghemoglobin.
 
NH4+ is assimilated into organic N before it can be exported.
NH4+ is incorporated into
a)  Amides (ASP-NH2) OR
b)  Uredes
Amides are exported from nodules via xylem.

AMIDES ARE CONVERTED TO
Amino acids, proteins or
Nucleic acids, like DNA, and RNA

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Lecture Review

1.  The development of a seed involves three events:
 |___(1)____|_________(2)__________|___(3 )____|
           0                30                                     100            120 days after flowering
 -(1)  embryo formation, -(2) reserve food accumulation, (3)  onset of dormancy
Mature soybean seeds contain: 37% protein,   17% lipid
     26% carbohydrate (starch)
Soybean seeds have 2-3X more protein than cereal grains.  Where does this protein come from?  Use the questions & table to guide your answer.
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                                                      A. Soybean/in NO3 containing soil         B. Soybean/in soil with
                                                                         No Rhizobium                       Rhizobium, but no NO3
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i.  N form taken up from environment             _______________             ___________
ii.  N form in (i) is too oxidized, so it is converted to __________             ___________

iii.  Enzyme(s) needed for conversion in (ii) _________________             ____________
      _________________ ____________
iv.  Is there a loss or gain of electrons (iii)? _________________             ____________
What essential substrate(s) is(are) needed? _________________             ____________
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v.  The first amino acid formed from (ii) is Asparagine.
 There are ____(number) C & ____(n) N in one molecule of asparagine.
The structure of asparagine (2 more pts for full pts):
 

vii)  What is the ultimate source of C in asparagine and thus all amino acids and proteins?
Explain.
 

(viii)  N from the environment is taken up by the _________ (organ), and  transported efficiently in the chemical form ________________ to the developing soybean seed via ______________ (tissue and organ).
 
 

x)  What is the function of the following and give a specific example:
-stored protein in the seed:
 

-stored starch in the seed
 

xi)  All the N atoms and C atoms in our bodies come from plants/algae.  True or False.
Briefly explain why for N and for C.