Roots and Root-Soil Relations
& Nitrogen Assimilation

Chp. 13 & 8

 

Roots, Soils, and Nutrient Uptake

With the exception of carbon and oxygen, which are supplied as carbon dioxide from the air, terrestrial plants generally take up nutrient elements from the soil solution through the root system.

The root systems of most plants are surprisingly extensive.

Through a combination of primary roots, secondary and tertiary branches, and root hairs, root systems penetrate massive volumes of soil in order to mine the soil for required nutrients and water.

 Soil is a complex medium.

 

It consists of a solid phase that includes mineral particles derived from parent rock plus organic material in various stages of decomposition, a liquid phase that includes water or the soil solution, gases in equilibrium with the atmosphere, and a variety of microorganisms.

 The solid phase, in particular the mineral particles, is the primary source of nutrient elements.

 In the process of weathering, various elements are released into the soil solution, which then becomes the immediate source of nutrients for uptake by the plant.

 

THE SOIL AS A NUTRIENT RESERVOIR

Soils vary widely with respect to composition, structure, and nutrient supply.

 Especially important from the nutritional perspective are inorganic and organic soil particles called colloids.

 Soil colloids retain nutrients for release into the soil solution where they are available for uptake by roots.

Thus, the soil colloids serve to maintain a reservoir of soluble nutrients in the soil.

 

In this chapter we will examine the availability of nutrients in the soil and their uptake by roots. This will include

• the soil as a source of nutrient elements, the colloidal nature of soil, and ion exchange properties that determine the availability of nutrient elements in a form that can be taken up by roots

 

• mechanisms of solute transport across membranes, including simple and facilitated diffusion and active transport, the function of membrane proteins as ion channels and carriers, and the role of electrochemical gradients

•        ion traffic into and through the root tissues and the concept of apparent free space and

•        the beneficial role of microorganisms, especially fungi, with respect to nutrient uptake by roots.


  Nutrient Uptake & Sustainable Agriculture

•     A deficiency of an element makes it difficult or impossible for the plant to complete a vegetative or reproductive stage of development

•     A deficiency can be prevented or corrected by supplying the element

 

Nutrient Uptake – from soil to plant via root

•       Movement to the roots: 

•        1) Root extension - exposure to soil and new supplies of nutrients - roots could contact 3% of the soil or nutrients in the soil.  

2) Mass Flow –

Ψ    water absorbed by the root creates a water deficit near the root,

Ψ    more water moves to the root carrying nutrients with the water. 

Ψ    Important for nutrients in large quantities in the soil solution - N, K & Ca

 

NUTRIENT UPTAKE BY PLANTS REQUIRES TRANSPORT OF THE NUTRIENT ACROSS ROOT CELL   MEMBRANES

In order for mineral nutrients to be taken up by a plant, they must enter the root by crossing the plasma membranes of root cells.

 From there they can be transported through the symplast to the interior of the root and eventually find their way into the rest of the plant.

 

•Nutrient uptake by roots is therefore fundamentally a cellular problem, governed by the rules of membrane, transport.

•There are, however, three fundamental concepts--simple diffusion, facilitated diffusion, and active transport-that have persevered, largely because they have proven particularly useful in categorizing and interpreting experimental observations.

Nutrient Uptake

3) Diffusion - movement of nutrients due to an imbalance of concentration ( diffusion gradient)

Requirements for nutrient uptake by plants

•     Actively growing plants - anything that affects the metabolism of the plant will affect nutrient uptake

•     Metabolic energy is required.   Plant roots must be able to respire. Soils must have oxygen

Conditions required for Nutrient Uptake by plants

 

Nutrient Uptake

Nutrient Uptake

Functions of Roots

•     Absorption

•     Anchorage

•     Storage

 

Root Systems

•     Tap vs. fibrous root systems

Root Systems

•     Roots may occupy about 1% of total soil volume (with actively growing crops)

•     In wildlands (e.g. forest, grassland), roots may represent up to 50% of total plant mass (roots + shoots)

•     In agriculture, roots may represent about 25% of total plant mass

Depth of Root Systems

•     Lettuce                     30 cm

•     Cotton                      120-150 cm

•     Alfalfa                      250-300 cm

•     Turf             60-90 cm

•     Mesquite       50 m ?????

•     Regardless of root system depth, the most important roots for nutrient and water uptake are usually found in the top 15-20 cm of soil.

Roots and Plant Survival Strategies

•     Desert Perennials (other than cacti)

–    Much of their biomass is below ground

•     Desert Annuals—small root systems

•     Phreatophytes (e.g. mesquite)

–    “Dual” Root system

•  Shallow for capturing rainfall

•  Deep for tapping into the water table

•     Many plant species, when confronted with drought or nutrient deficiency, will increase root growth at the expense of shoots.

Root Morphology

•     Longitudinal

–    Meristematic zone

–    Elongation zone

–    Maturation zone

–    Mature zone

•     Cross-sectional

–    Epidermis

–    Cortex

–    Endodermis

–    Stele

Root Morphology

 

Functions

•     Root cap

–    Protection of meristem

–    Secretion of mucigel

–    Initiation of symbiotic relationships

•     Elongation Zone

–    Elongation of cells forces root through the soil

•     Maturation zone

–    Root Hairs

–    Major zone of water and nutrient uptake

 

 

 

 

 

 

 

 

 

Root Hairs

•     Single-cell extensions of epidermal cells

•     Tremendously important for providing surface area for water and nutrient uptake

•     Produced in the maturation zone

•     Sites of infection by pathogens, N fixing bacteria, and mycorrhizae

Root Hairs

•     Root hairs can account for 2/3 of total root surface area.







•     Root hairs are fragile and susceptible to breakage as soils dry.

Size of Roots

•     Fine lateral roots are 0.1 to 0.2 mm in diameter.

 

•     Root hairs are 0.01 to 0.05 mm in diameter.

 

•     Soil “micropores” are considered to be those <0.08 mm in diameter.

 

•     Therefore, root hairs are important for accessing water and nutrients in micropores.

Mature Roots

•     The root epidermis and endodermis become covered with a waxy substance known as “suberin”.

 

•     Function:
Water and nutrient uptake rates are lower in mature root zones than in immature:

–    Lower permeability

–    Formation of aerenchyma (air pockets)

 

Aerenchyma

Implications of Root Morphology

•     The youngest part of the root is more permeable to water and nutrients than is the older part (behind the maturation zone).

 

•     Most nutrients are taken up in  the younger root zone (‘feeder roots’) with abundant root hairs.

 

•     Excessive soil drying and mechanical disturbance will damage young roots, reduce nutrient uptake.

The Rhizosphere

•     Definition:  a zone of soil where microbial         activity is influenced by roots

•     Why?  Border cells, root leakage and     secretion of organic compounds

•     May extend to about 2 mm from root surface.

The Rhizosphere

Properties:

–    Higher available C for microbial growth

–    Higher microbial population and activity

–    Lower pH than “bulk” soil

–    Lower O2 than “bulk” soil

–    Altered nutrient availability for plants

Border Cells in the Rhizosphere

Organic C in the Rhizosphere (Rhizodeposition)

•     Plants release simple and complex carbohydrates, nucleic acids, enzymes into the rhizosphere

•     These compounds are used by microorganisms as sources of C and energy.

•     Up to 30% of plant C fixed in photosynthesis may be released from roots.

Importance of the Rhizosphere

•     A healthy rhizosphere will help plants by:

–    Increasing nutrient availability

–    Suppressing pathogens

–    Increasing water availability

•     However, the effects of agricultural management on the rhizosphere are still largely unknown.

 

 

Mycorrhizae (my·cor·rhi·za \ˌmī-kə-ˈrī-zə) were originally discovered by the nineteenth-century German botanist A. B. Frank, who concluded, on the basis of experiments conducted with beech seedlings, that mycorrhizal inoculation stimulated seedling growth.

Although not universally accepted in the beginning, these results have been amply confirmed by more modern studies.

Numerous studies with pine and other tree seedlings in the United States, Australia, and the former Soviet Union have demonstrated 30 to 150 percent increases in dry weight of tree seedlings infected with mycorrhizae when compared with noninfected controls.

 

. The primary cause of mycorrhizal-enhanced growth appears to be enhanced uptake of nutrients, especially phosphorous.

In a classic experiment, Hatch demonstrated in 1937 that infected pine seedlings absorbed two to three times more nitrogen, potassium, and phosphorous.

 Coupled with enhanced nutrient uptake is the observation that mycorrhiza-induced growth responses are more pronounced in nutrient-deficient soils.

Mycorrhizae

•     Two kinds:

–    Ectomycorrhizae


–    Endomycorrhizae

 

 

MYCORRHIZAE ARE FUNGI THAT INCREASE THE VOLUME OF THE NUTRIENT DEPLETION ZONE AROUND ROOTS

•            Perhaps the most widespread-and from the nutritional perspective, more significant-associations between plants and microorganisms are those formed between roots and a wide variety of soil fungi.

•            A root infected with a fungus is called a mycorrhiza (literally, fungus root).

•            Mycorrhizae are a form of mutualism, an association in which both partners derive benefit.

•            The significance of mycorrhizae is reflected in the observation that more than 80 percent of plants studied, including virtually all plant species of economic importance, form mycorrhizal associations.

•             

 

•Two major forms of mycorrhizae are known: ectotrophic and endotrophic

 

•The ectotrophic form, also known as ectomycorrhizae, is restricted to a few families consisting largely of temperate trees and shrubs,the intercellular or apoplastic space of the root cortex,such as pines (Pinaceae) and beech (Fagaceae).

• Ecto-forming an intercellular network called a Hartig net-mycorrhizae are typically short, highly branched.     

 

•Endotrophic mycorrhizae, or endomyensheathed by a tightly interwoven mantle of fungal mycorrhizae, are found in some species of angiosperm family and most gymnosperms as well (except the Pinaceae).

•Unlike the ectomycorrhizae, the hyphae of endomycorrhizae develop extensively within cortical cells of the host roots.

 

•     Plants can also increase effective root length/surface area by  investing in symbioses with soil fungi, mycorrhizae…

•     Mycorrhizae, by increasing effective root length, increase the  volume of soil exploited by roots

•     Most plants, (around 75%), are mycorrhizalIn mutualistic mode, the plant provides photosynthate (fixed C) to the fungus, whereas the fungus provides nutrients and water to the  plant

•     The ‘mycorrhiza’ (fungus-root) is the interface where fungal hyphae actually penetrate the root, forming an interface where nutrients and carbon are exchanged

•     The structure and nature of this interface varies somewhat among mycorrhizal types

 

There are four major types of mycorrhizae:

1. ectomycorrhizae: temperate, boreal, some tropical forests

2. arbuscular mycorrhizae (AM, used to be called VAM): herbaceous communities and tropical forests

3. ericoid mycorrhizae: heathlands, tundra

4. orchid mycorrhizae

ecto- and arbuscular- mycorrhizae are the most widespread

 

Arbuscular Mycorrhizae

– fungal hyphae penetrate walls of cortical cells (but not plasma membrane)

– produce highly branched "arbuscules" in close association with plant cell plasma membrane, forming the point of transfer of nutrients and carbon

– especially important for P uptake

– also important in water stress

 

 

 

 

 

•     Ectomycorrhizae

•     common in woody plants

•      Mantle or sheath of hyphae surrounds the root

•     Hyphae penetrate intercellular spaces of root cortex to form  Hartig Net (point of material exchange between plant and fungus),  but do not penetrate cortical cell walls

•     plants respond to infection by growing short branched "club" roots

•     important in acquiring P and N; fungus produces proteases  that cleave proteins into amino acids

•     may be important in water stress

 

 

Roots and Mycorrhiza – an old symbiosis

•     Mutual benefit

•      Carbohydrates for the fungus

•      P, Zn, Cu, water, N for plant

 

•     Different types

•     1. Vesicular-arbuscular mycorrhiza – VA-mycorrhiza

•     2. Ectomycorrhiza

•     Other types

•             ericoid

•             orchid endomycorrhiza

 

 

 

•     1. Vesicular arbuscular mycorrhiza (AM)

 

•      Glomales (130 species – infects 300.000 plant species)

 

•      Found on roots of herbaceous angiosperms, most trees, mosses, ferns…

 

•      not present on Cruciferae, Chenopodiaceae, Proteaceae

 

•      small biomass compared to roots

 

 

•      Vesicular Arbuscular Mycorrhiza

 

•      Inside root

•       Intercellular mycelium

•       Intracellular arbuscule

–     tree-like haustorium

•       Vesicle with reserves

•      Outside root

•       Spores (multinucleate)

•       Hyphae

–    thick runners

–    filamentous hyphae

•      Form extensive network of hyphae

•      even connecting different plants

 

 

 

 

 

 

 

 

 

 

 

 

•     Summary on mycorrhizae

 

•      Symbiosis with mycorrhiza allows greater soil exploration,

•       and increases uptake of nutrients (P, Zn, Cu, N, water)

 

•      Great SA per mass for hyphae vs. roots

 

•      Mycorrhiza gets carbon from plant

 

•      Two main groups of mycorrhiza – Ectomycorrhiza and VA-mycorrhiza

 

•     Ectomycorrhizae

•     common in woody plants

•     Mantle or sheath of hyphae surrounds the root

•     Hyphae penetrate intercellular spaces of root cortex to form Hartig Net (point of material exchange between plant and fungus),  but do not penetrate cortical cell walls

•     plants respond to infection by growing short branched "club" roots

•     important in acquiring P and N; fungus produces proteases
that cleave proteins into amino acids

•     may be important in water stress

 

 

 

 

 

 

 

 

•     Two Other Important Types of Mycorrhizae

•     Ericoid mycorrhizae (ericoid – fungus associations)
  – like EM fungi, ericoid fungi produce proteases (enzymes that hydrolyze organic forms of N); amino acids can then be transferred to the host plant

•     – thus, important in plant N nutrition

•     – ericoid fungi also produce phenol oxidases, capable of humus  degradation

•     – ericoid hyphae do not extend as far beyond the root as EM  fungi

•     Orchid mycorrhizae (orchid – fungus associations)

•     – in some cases, fungus provides carbon as well as nutrients!)

 

Mycorrhizae

Mycorrhizae

Mycorrhizae

•     A fungal-root symbiosis

–    increase root length and surface area

–    help plants to take up nutrients that are “immobile” in soil (e.g. P)

–    inhibit heavy metal uptake

–    increase water uptake by roots

–    are vital for the growth and survival of plants where nutrients are likely to be limiting, and/or where heavy metals are problems

 

Managing Mycorrhizae

•     About 80% of all plants have mycorrhizael associations.

•     Innoculation (addition) with mycorrhizae is often used for nursery and forestry plantings.

•     Addition of mycorrhizae to agricultural soils has shown few benefits so far:

–    Difficulty of establishing Endo fungi

Nitrogen Fixation

•     Definition:  The conversion of atmospheric N2 to forms usable by plants.

•     Certain bacteria and actinomycetes can carry out N fixation.

•     The most important N fixation occurs through a symbiotic relationship between legume roots and bacteria.

Legume Root Nodules

Nitrogen Fixation

•     Legumes commonly form N-fixation symbioses

•     N fixation is not free

–    may cost the plant up to 30% of the C it captures from the atmosphere in photosynthesis

•     Rate of N fixation is inversely proportional to available N in the soil.

Importance of N Fixation

•     Legume crops can derive virtually all their N requirement through N fixation.

 

•     In crop rotations, legume crop residues can be incorporated into the soil. Upon decomposition, N is released.

 

•     Legumes and non-legumes can be grown together—in this case 20-50% of the N fixed in the legume may become available to the non-legume.

Final Thoughts on Roots

•     Root system is responsible for water and nutrient uptake.

•     The youngest part of the root system is responsible for much of this uptake

–    Root hairs are critical, fragile and can be easily damaged by compaction, tillage, etc.

•     The root system is dynamic and will respond to changes in soil conditions.

 

Final Thoughts (2)

•     The root is part of a microbial community known as the rhizosphere

•     A healthy rhizosphere is important for plant health

–    There’s a lot about it we don’t know yet

•     Mycorrhizae - water, nutrients, pathogen protection

•     Rhizobia - N fixation

Assimilation and fixation of nitrogen

 

 

Remember, Plants are:

•     Capable of making all necessary organic compounds from inorganic compounds and elements in the environment (autotrophic)

•     Required to compete with other organisms for these nutrients

•     Required to employ complex energetic pathways to convert macronutrients to useable forms

Nitrogen in the environment

•     Many biochemical compounds present in plant cells contain nitrogen

–    Nucleoside phosphates

–    Amino acids

•     These form the building blocks of nucleic acids and protein respectively

 

•     Only carbon, hydrogen, and oxygen are nor abundant in plants than nitrogen

 

 

•     Nitrogen Assimilation

•     On a dry-weight basis, nitrogen is the fourth most abundant nutrient element in plants.

•      It is an essential constituent of proteins, nucleic acids, hormones, chlorophyll, and a variety of other important primary and secondary plant constituents.

•      Most plants obtain the bulk of their nitrogen from the soil in the form of either nitrate (NO3) or ammonium (NH4), but the supply of nitrogen in the soil pool is limited and plants must compete with a variety of soil microorganisms for what nitrogen is available.

•      As a result, nitrogen is often a limiting nutrient for plants, in both natural and agricultural ecosystems.

Nitrogen in the environment

•     Present in many forms

•     78% of atmosphere is N2

–    Most of this is NOT available to living organisms

•     Getting N2 for the atmosphere requires breaking the triple bond between N2 gas to produce:

•     Ammonia (NH3)

•     Nitrate (NO3-)

•     So, N2 has to be fixed from the atmosphere so plants can use it

 

Nitrogen in the environment

•     This occurs naturally by:-Lightning:

–    8%: splits H2O: the free O and H attack N2 – forms HNO3 (nitric acid) which fall to ground with rain

•     Photochemical reactions:

–    2%: photochemical reactions between NO gas and O3 to give HNO3

•     Nitrogen fixation:

–    90%: biological – bacteria fix N2 to ammonium (NH4+)

 

Nitrogen in the environment

 

•     Once fixed in ammonium or nitrate :-

–    N2 enters biochemical cycle

–    Passes through several organic or inorganic forms before it returns to molecular nitrogen

–    The ammonium (NH4+) and nitrate (NO3-) ions generated via fixation are the object of fierce competition between plants and microorganisms

–    Plants have developed ways to get these from the soil as fast as possible

Root uptake soon depletes nutrients near the roots

•     Formation of a nutrient depletion zone in the region of the soil near the plant root

–    Forms when rate of nutrient uptake exceeds rate of replacement in soil by diffusion in the water column

–    Root associations with Mycorrhizal fungi help the plant overcome this problem

Mycorrhizal associations

•      Not unusual

–     83% of dicots, 79% of monocots and all gymnosperms

•      Ectotrophic Mycorrhizal fungi

–     Form a thick sheath around root.  Some mycelium  penetrates the cortex cells of the root

–     Root cortex cells are not penetrated, surrounded by a zone of hyphae called Hartig net

–     The capacity of the root system to absorb nutrients improved by this association – the fungal hyphae are finer than root hairs and can reach beyond nutrient-depleted zones in the soil near the root

Mycorrhizal associations

Nutrients move from fungi to root cells

•     Ectotrophic Mycorrhizal

–    Occurs by simple diffusion from the hyphae in the hartig net to the root cells

•     Vesicular arbuscular mycorrhizal fungi

–    Occurs by simple diffusion from the arbuscules to the root cells

–    Also, as arbuscules are degenerating as new ones are forming, the nutrients may be released directly into the host cell

Stored ammonium can be toxic

•     Plants can store high levels of nitrate or translocate it via the phloem without any effect.

•     However, high levels of ammonium are toxic

–    Dissipates transmembrane proton gradients required for both photosynthetic and respiratory electron transport

–    AND movement of metabolites to vacuoles.

 

Stored ammonium can be toxic

•     At high pH in stroma, matrix or cytoplasm:

–    Ammonium reacts with OH- to produce NH3.  

•     NH3 is membrane permeable and diffuses freely across a membrane down a concentration gradient

•     At low pH in intermembrane space, lumen, or vacuole:

–    NH3 reacts with H+ to form ammonium

 

Remember: Nitrogen – the most important mineral nutrient in the soil

•     Nitrogen is frequently limiting in in terrestrial systems terrestrial systems

 

•     Microbial activity is continually converting N to lower energy forms

 

•     Conversion to organic form requires raising N to higher energy levels

Nitrate Assimilation

Nitrogen assimilation

 

            NO3      NO2       NH4+       amino acids

    nitrate        nitrite      ammonium

 

•     Requires large input of energy

•     Forms toxic intermediates

•     Mediated by specialized enzymes that are closely regulated are closely regulated

•     Doesn’t have to start at the beginning

Nitrogen assimilation

•     Plants assimilate most of the nitrate absorbed by their roots into organic nitrogen compounds.

•     The first step of this process is the reduction of nitrate to nitrite in the cytosol by the enzyme nitrate reductase.

 

 

Nitrogen assimilation

NAD(P)H induces NADH or NADPH

–    The most common form of nitrate reductase uses only NADH as an electron donor

–    The nitrate reductases of higher plants are composed of two identical sub-units, each containing three prosthetic groups

•   FAD—flavin adenine dinucleotide

•   Heme

•   Molybdenum—organic molecule called pterin

Nitrate Assimilation

•     Nitrate reductase is the main molybdenum containing protein in vegetative tissues

•     Nitrate levels, light intensity, and concentration of carbohydrates all influence the activity of nitrate reductases at the transcription and translation levels

–    These factors stimulate a protein, phosphatase, that dephosphorylates several serine residues on the nitrate reductase protein thereby activating the enzyme

 

–    This dephosphorylation/phosphorylation cycle provides more rapid control over this enzyme than degredation/synthesis of new enzyme would achieve ( minutes versus hours)

Nitrite Reductase Converts Nitrite to Ammonium

•     Nitrite (NO2-)is highly reactive

•     Plant cells immediately transport the nitrite generated by nitrite reduction from the cytosol into chloroplasts in leaves and plastids in roots

–    In these organelles, nitrite reductase reduces nitrite to ammonium

 

Nitrite Reductase Converts Nitrite to Ammonium

•     Chloroplast and root plastids contain different forms of the enzyme, but both forms consist of a single polypeptide containing an iron sulfur cluster and a specialized heme group

–    The heme does redox reactions and electron flow, just like the reaction sites of chlorophyll

 

Nitrite Reductase Converts Nitrite to Ammonium

•     Nitrite is highly reactive

•     Plant cells immediately transport the nitrite generated by nitrite reduction from the cytosol into chloroplasts in leaves and plastids in roots

–    In these organelles, nitrite reductase reduces nitrite to ammonium

•     Chloroplast and root plastids contain different forms of the enzyme, but both forms consist of a single polypeptide containing an iron sulfur cluster and a specialized heme group

–    The heme does redox reactions and electron flow, just like the reaction sites of chlorophyll

Plants assimilate nitrate in both roots and shoots

•     In many plants, when the roots receive small amounts of nitrate, this nitrate is reduced primarily in the roots

 

•     As nitrate supply increases, a greater proportion of the absorbed nitrate is translocated to the shoot and assimilated there

 

•     Generally, species native to temperate rely more heavily on nitrate assimilation by the roots than do species of tropical or subtropical origins

Ammonium Assimilation

•     Plants cells avoid ammonium toxicity by rapidly converting the ammonium generated from nitrate assimilation or photorespiration into amino acids

•     This requires the action of two enzymes

–    Glutamine synthetase – combines ammonium with glutamate to form glutamine

–    Glutamate synthase – stimulated by elevated levels of glutamine synthetase

–    Transfers the amino group of glutamine to an intermediate yielding two molecules of glutamate

 

•     This accounts for most of the fixation of atmospheric N2 into ammonium

–    Represents the key entry point of molecular nitrogen into the biogeochemical cycle of nitrogen

•     Free living and symbiotic bacteria are responsible for converting atmospheric nitrogen into ammonium

•     Most of these are free living in the soil, a few form symbiotic associations with higher plants

–    The prokaryote directly provides the host plant with nitrogen in exchange for other nutrients and carbohydrates

•     The most common association is between members of the plant family leguminosae and bacteria of the genera Azorhizobium

 

Nitrogen Fixation Requires Anaerobic Conditions

•     As oxygen irreversibly inactivates the nitrogenase enzymes involved in nitrogen fixation, nitrogen must be fixed under anaerobic conditions

 

–    Therefore each of the nitrogen-fixing organisms either functions under natural anaerobic conditions or can create an internal anaerobic environment in the presence of oxygen

Nitrogen Fixation Requires Anaerobic Conditions

•     In cyanobacteria, anaerobic conditions are created in specialized cells called heterocysts

–    These are thick-walled cells which lack photosystem II—the oxygen producing photosystem of chloroplasts

•     Cyanobacteria can fix nitrogen under anarobic conditions such as those that occur in flooded fields

–    In Asian countries, nitrogen fixing cyanobacteria of both the heterocyst and non-heterocyst types are the major means of maintaining an adequate nitrogen supply in rice fields

•   They fix nitrogen when the fields are flooded, and die as the fields dry, releasing the fixed nitrogen into the soil

 

Symbiotic Nitrogen Fixation Occurs in Specialized Structures

•     Symbiotic nitrogen-fixing prokaryotes dwell within nodules

–    Special organs of the plant host that enclose the nitrogen-fixing bacteria

•     Grasses can also develop symbiotic relationships with nitrogen-fixing organisms, but these associations do not lead to the formation of root nodules

–    Nitrogen-fixing bacteria seem to colonize plant tissues or anchor to the root surface, mainly around the elongation zone and the root hairs

•   Known as actinorhizal plants

 

Symbiotic Nitrogen Fixation Occurs in Specialized Structures

•      Both legumes and actinorhizal plants regulated gas permeability in their root nodules

–    Maintaining a level of oxygen within the nodule that can support cellular respiration for the bacteria, but still sufficiently low to avoid inactivation of the nitrogenase

Nodule formation involves several phytohormones

•      During root nodule formation, two process occur simultaneously

•      Infection and Nodule Organogenesis

–    (A) Rhizobia attach to the root hairs and release nod factors that produce a pronounced curling of the root hair cell

–    (B) Rhizobia get caught and curl, degrade the root hair cell wall allowing the bacterial cells direct access to the outer surface of the plant plasma membrane

Nodule formation involves several phytohormones

•      (C) Then the infection thread forms

–    Formed from Golgi depositing material at the tip at the site of infection.  Local degradation of root hair cell wall also occurs

•      (D) Infection thread reaches the end of the cell, and thread plasma membrane fuses with plasma membrane of root hair cell

–    Then bacterial cells are released into the fused plasma membranes

Nodule formation involves several phytohormones

•      (E) Rhizobia are released into the apoplast and enter the middle lamella,

–    This leads to the formation of a new infection thread, which forms an open channel with the first

•      (F) Infection thread expands and branches until it reaches target cells

–    Vesicles composed of plant membrane enclose bacterial cells and they are released into the cytoplasm

Nodule formation involves several phytohormones

•     At first bacteria continue to grow with vesicles expanding by fusing with smaller vesicles

•     Following an as yet determined chemical signal from the plant, bacteria stop dividing and differentiate

–    Forms nitrogen-fixing organelles called bacteroids

•     The nodule itself develops a vascular system

–    To exchange fixed nitrogen for nutrients from the plant

•     And a layer of cells to exclude O2 from the rood nodule interior

The nitrogenase enzyme complex fixes N2

•     Biological nitrogen fixation produces ammonium (NH3) from molecular nitrogen.

 

•       N2 + 8e- + 8H+ + 16 ATP        2NH3 + H2+ 16 ADP + 16 Pi

 

•       Note that the reduction of N2 to 2NH3 is a six-electron transfer, and is coupled to the reduction of two protons to evolve H2

•     This reaction is catalyzed by nitrogenase enzyme complex

The nitrogenase enzyme complex fixes N2

•     Can be separated into two components

–    The Fe protein

–    The MoFe protein

•     Neither of which has catalytic activity by itself

The nitrogenase enzyme complex fixes N2

•     Ferredoxin reduces the Fe protein

–    Binding and hydrolysis of ATP to the Fe protein is thought to cause a conformational change of the Fe protein that facilitates the REDOX reactions

•       The Fe protein reduces the MoFe protein, and the MoFe protein reduces the N2

The MoFe protein can reduce many substances 

•     The MoFe protein can reduce many substrates

•     Although under natural conditions the MoFe only reacts with N2 and H+.

Summary

•     Nutrient assimilation is the process by which nutrients acquired by plants are incorporated into the carbon constituents necessary for growth and development.

•     For Nitrogen:

•     Assimilation is but one in a series of steps that constitute the nitrogen cycle.

•     The principal sources of nitrogen available to plants are nitrate (NO3-) and ammonia (NH4+). 

•     Nitrate absorbed by roots is assimilated in either shoots or roots

–    depending on nitrate availability and plant species

 

•     In nitrate assimilation, nitrate (NO3-) is reduced to nitrite (NO2-) in the cytosol via the enzyme nitrate reductase.

•     Then nitrite is reduced to ammonium (NH4+) in roots by nitrite reductase.

•     Ammonium (NH4+) from either root absorption or generated through nitrate assimilation  or photorespiration is converted glutamine or glutamate through the sequential actions of glutamine synthase and glutamate synthase.

•     Once assimilated into either glutamine or glutamate, nitrogen mat be transferred to many other organic compounds

–    Via transaminatation reactions

 

Summary

•     Many plants form a symbiotic relationship with nitrogen fixing bacteria that contain an enzyme complex, nitrogenase, that can reduce atmospheric nitrogen to ammonia.

•     Legumes and actinorhizal plants form associations with rhizobia.

•     These associations result from a finely tuned interaction between the bacteria and the host plant

–    Involves the recognition of specific signals between the symbiotic bacteria and the host plant

Any Questions?

 

 

•            Nitrogen - the most limiting soil nutrient

•            Evidence - factorial fertilization experiments (N, P, K, etc.)show largest growth response to N.

•            Required in greatest amount of all soil nutrients

•            2. A component of proteins (enzymes, structural proteins,

•            chlorophyll, nucleic acids)

•            3. The primary photosynthetic enzyme, Rubisco, accounts for a 25 to 50% of leaf N.

 

•                Photosynthetic capacity is strongly correlated with  leaf N concentration.

•            4. Availability in most soils is low

•            5. Plants spend a lot of energy on N acquisition – growing roots, supporting symbionts, uptake into roots, biochemical assimilation into amino acids, etc.  

 

 

•            The inorganic forms of nitrogen in soils.

 

•            NH4+, ammonium ion. A cation that is bound to clays.

•            NO3-, nitrate ion. An anion that is not bound to clays.

•            Nutrient “mobility” in soils refers to the rate of diffusion, which is influenced by nutrient ion interactions with soil particles.

•            Would you expect NH4+ or NO3-  to diffuse more rapidly?

•            Would you expect a more pronounced depletion zone for NH4+ or NO3-?

 

 

 

 

 

 

 

 

•The beneficial role of mycorrhizae, particularly with respect to the uptake of phosphorous, appears to be related to the nutrient depletion zone that surrounds the root .

•This zone defines the limits of the soil from which the root is able to readily extract nutrient elements.

•Additional nutrients can be made available only by extension of the root into new regions of the soil or by diffusion of nutrients from the bulk soil into the depletion zone.

•The extent of the depletion zone varies from one nutrient element to another, depending on the solubility and mobility of the element in the soil solution.

 

•The depletion zone for nitrogen, for example, extends some distance from the root because nitrate is readily soluble and highly mobile. Phosphorous, on the other hand, is less soluble and relatively immobile in soils and, consequently, the depletion zone for phosphorous is correspondingly smaller.

•Mycorrhizal fungi assist in the uptake of phosphorous by extending their mycelia beyond the phosphorous depletion zone.

• Apparently, mycorrhizal plants find it advantageous to expend their carbon resources supporting mycorrhizal growth as opposed to more extensive growth of the root system itself.

 

As we continue to learn about mycorrhizae, their nutritional role becomes increasingly evident.

Many mycorrhizae are host species-specific.

Attempts to establish a plant species in a new environment may be unsuccessful if the appropriate mycorrhizal fungus is not present.

Inoculation of fields with mycorrhizal fungi is now an additional factor taken into account by the forest and agricultural industries when attempting to resolve problems of soil infertility.

 

CHAPTER REVIEW

1. Distinguish between simple diffusion, facilitated diffusion, and active transport. Which of these three mechanisms would most probably account for:

(a) entry of a small lipid-soluble solute;

(b) extrusion of sodium ions leaked into a cell; (c) rapid entry of a neutral hydrophilic sugar; (d) accumulation of potassium ions?

2. The Casparian band was encountered earlier with regard to root pressure and again in this chapter with regard to ion uptake. What is the Casparian band and how does it produce these effects?

3. Trace the pathway taken by a potassium ion from the point where it enters the root to a leaf epidermal cell.

4. What are channel proteins and what role do they play in nutrient uptake?

5. Describe the concept of apparent free space. What role does apparent free space play in the uptake of nutrient ions?

6. Why is an accumulation ratio greater than 1.0 not necessarily an indication that active transport is involved?